LABORATORY DIAGNOSTIC OF INVASIVE FUNGAL INFECTIONS

Mardiastuti H Wahid Department of Microbiology

Faculty of Medicine Universitas Indonesia

INTRODUCTION

Increased incidence of IFI

due to increased immuno-

compromised condition

malignancies

Increased resistance

to antimycotic

agents

The need for

accurate laboratory diagnostics

Better treatment and clinical outcome

LABORATORY DIAGNOSTIC OF IFI

bull (13)-β-D-Glucan assay accurate laboratory diagnostic tool in determining between definitive and probable invasive fungal infections

bull May be useful in the clinical practice if

ndash Implemented in a proper setting

ndash Interpreted after consideration of its limitation

Drosos E Karageorgopoulos et al Clinical Infectious Diseases 201152(6)750ndash770

bull The Glucatell serum BG detection assay is highly sensitive and specific as a diagnostic adjunct for IFI

Zekaver Odabasi et al

Clinical Infectious Diseases 2004 39199ndash205

LABORATORY DIAGNOSTIC OF IFI

INVASIVE CANDIDOSIS

SPECIES DISTRIBUTION

bull The incidence of Candidosis infection in hospitalized patients in the USA has increased (1980-1989) 8 responsible for all bloodstream infections

bull In the 1990s incidence of Candida bloodstream infections decreased due to a decline of C albicans infections however the incidence rate of C glabrata increased

bull 1992-1993 1998-2000 overall incidence of Candida bloodstream infections increased

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

bull Invasive candidosis

ndash endogenous

ndash transmission from person to person (direct patient-patient transmission healthcare workers medical devices contaminated intravenous infusions)

bull Cross infection in ICU

ndash C albicans via hand of healthcare workers

ndash C parapsilosis via medical devices

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

bull Invasive candidosis

ndash Neutropenic cancer patients

ndash Stem cellorgan transplants recipients

ndash ICU patients (adult surgical neonatal)

Widespread use of fluconazole emergence of less susceptible species

Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results

bull Dignosis based on clinical radiological microbiological and histopathological findings

bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Investigation included

ndash Microscopy

ndash Culture

ndash Serological test

ndash D-arabinitol detection

ndash Molecular assays

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids

bull C glabrata produces only yeast cells and

C albicans produces true hyphae in tissues

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection

bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites

bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication

bull Different susceptibility among Candida sp completed identification (speciation) before treatment

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis

bull Isolation of Candida sp from urine may indicate a serious infection

bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis

bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Limited usefulness

bull False positive (mucosal colonization superficial infection)

bull False negative (immunocompromised)

bull Antigen detection mannan insensitive

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

DETECTION OF D-ARABINITOL

bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)

bull It can be detected in serum or urine

bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR ASSAY

bull PCR detection of Candida DNA in blood serum LCS other specimens

bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid

bull False positive

bull Standardized commercial assays were not available

bull Can not be recommended in diagnosis of invasive candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

INTRODUCTION

Increased incidence of IFI

due to increased immuno-

compromised condition

malignancies

Increased resistance

to antimycotic

agents

The need for

accurate laboratory diagnostics

Better treatment and clinical outcome

LABORATORY DIAGNOSTIC OF IFI

bull (13)-β-D-Glucan assay accurate laboratory diagnostic tool in determining between definitive and probable invasive fungal infections

bull May be useful in the clinical practice if

ndash Implemented in a proper setting

ndash Interpreted after consideration of its limitation

Drosos E Karageorgopoulos et al Clinical Infectious Diseases 201152(6)750ndash770

bull The Glucatell serum BG detection assay is highly sensitive and specific as a diagnostic adjunct for IFI

Zekaver Odabasi et al

Clinical Infectious Diseases 2004 39199ndash205

LABORATORY DIAGNOSTIC OF IFI

INVASIVE CANDIDOSIS

SPECIES DISTRIBUTION

bull The incidence of Candidosis infection in hospitalized patients in the USA has increased (1980-1989) 8 responsible for all bloodstream infections

bull In the 1990s incidence of Candida bloodstream infections decreased due to a decline of C albicans infections however the incidence rate of C glabrata increased

bull 1992-1993 1998-2000 overall incidence of Candida bloodstream infections increased

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

bull Invasive candidosis

ndash endogenous

ndash transmission from person to person (direct patient-patient transmission healthcare workers medical devices contaminated intravenous infusions)

bull Cross infection in ICU

ndash C albicans via hand of healthcare workers

ndash C parapsilosis via medical devices

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

bull Invasive candidosis

ndash Neutropenic cancer patients

ndash Stem cellorgan transplants recipients

ndash ICU patients (adult surgical neonatal)

Widespread use of fluconazole emergence of less susceptible species

Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results

bull Dignosis based on clinical radiological microbiological and histopathological findings

bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Investigation included

ndash Microscopy

ndash Culture

ndash Serological test

ndash D-arabinitol detection

ndash Molecular assays

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids

bull C glabrata produces only yeast cells and

C albicans produces true hyphae in tissues

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection

bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites

bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication

bull Different susceptibility among Candida sp completed identification (speciation) before treatment

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis

bull Isolation of Candida sp from urine may indicate a serious infection

bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis

bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Limited usefulness

bull False positive (mucosal colonization superficial infection)

bull False negative (immunocompromised)

bull Antigen detection mannan insensitive

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

DETECTION OF D-ARABINITOL

bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)

bull It can be detected in serum or urine

bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR ASSAY

bull PCR detection of Candida DNA in blood serum LCS other specimens

bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid

bull False positive

bull Standardized commercial assays were not available

bull Can not be recommended in diagnosis of invasive candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

Increased incidence of IFI

due to increased immuno-

compromised condition

malignancies

Increased resistance

to antimycotic

agents

The need for

accurate laboratory diagnostics

Better treatment and clinical outcome

LABORATORY DIAGNOSTIC OF IFI

bull (13)-β-D-Glucan assay accurate laboratory diagnostic tool in determining between definitive and probable invasive fungal infections

bull May be useful in the clinical practice if

ndash Implemented in a proper setting

ndash Interpreted after consideration of its limitation

Drosos E Karageorgopoulos et al Clinical Infectious Diseases 201152(6)750ndash770

bull The Glucatell serum BG detection assay is highly sensitive and specific as a diagnostic adjunct for IFI

Zekaver Odabasi et al

Clinical Infectious Diseases 2004 39199ndash205

LABORATORY DIAGNOSTIC OF IFI

INVASIVE CANDIDOSIS

SPECIES DISTRIBUTION

bull The incidence of Candidosis infection in hospitalized patients in the USA has increased (1980-1989) 8 responsible for all bloodstream infections

bull In the 1990s incidence of Candida bloodstream infections decreased due to a decline of C albicans infections however the incidence rate of C glabrata increased

bull 1992-1993 1998-2000 overall incidence of Candida bloodstream infections increased

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

bull Invasive candidosis

ndash endogenous

ndash transmission from person to person (direct patient-patient transmission healthcare workers medical devices contaminated intravenous infusions)

bull Cross infection in ICU

ndash C albicans via hand of healthcare workers

ndash C parapsilosis via medical devices

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

bull Invasive candidosis

ndash Neutropenic cancer patients

ndash Stem cellorgan transplants recipients

ndash ICU patients (adult surgical neonatal)

Widespread use of fluconazole emergence of less susceptible species

Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results

bull Dignosis based on clinical radiological microbiological and histopathological findings

bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Investigation included

ndash Microscopy

ndash Culture

ndash Serological test

ndash D-arabinitol detection

ndash Molecular assays

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids

bull C glabrata produces only yeast cells and

C albicans produces true hyphae in tissues

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection

bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites

bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication

bull Different susceptibility among Candida sp completed identification (speciation) before treatment

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis

bull Isolation of Candida sp from urine may indicate a serious infection

bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis

bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Limited usefulness

bull False positive (mucosal colonization superficial infection)

bull False negative (immunocompromised)

bull Antigen detection mannan insensitive

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

DETECTION OF D-ARABINITOL

bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)

bull It can be detected in serum or urine

bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR ASSAY

bull PCR detection of Candida DNA in blood serum LCS other specimens

bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid

bull False positive

bull Standardized commercial assays were not available

bull Can not be recommended in diagnosis of invasive candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

LABORATORY DIAGNOSTIC OF IFI

bull (13)-β-D-Glucan assay accurate laboratory diagnostic tool in determining between definitive and probable invasive fungal infections

bull May be useful in the clinical practice if

ndash Implemented in a proper setting

ndash Interpreted after consideration of its limitation

Drosos E Karageorgopoulos et al Clinical Infectious Diseases 201152(6)750ndash770

bull The Glucatell serum BG detection assay is highly sensitive and specific as a diagnostic adjunct for IFI

Zekaver Odabasi et al

Clinical Infectious Diseases 2004 39199ndash205

LABORATORY DIAGNOSTIC OF IFI

INVASIVE CANDIDOSIS

SPECIES DISTRIBUTION

bull The incidence of Candidosis infection in hospitalized patients in the USA has increased (1980-1989) 8 responsible for all bloodstream infections

bull In the 1990s incidence of Candida bloodstream infections decreased due to a decline of C albicans infections however the incidence rate of C glabrata increased

bull 1992-1993 1998-2000 overall incidence of Candida bloodstream infections increased

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

bull Invasive candidosis

ndash endogenous

ndash transmission from person to person (direct patient-patient transmission healthcare workers medical devices contaminated intravenous infusions)

bull Cross infection in ICU

ndash C albicans via hand of healthcare workers

ndash C parapsilosis via medical devices

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

bull Invasive candidosis

ndash Neutropenic cancer patients

ndash Stem cellorgan transplants recipients

ndash ICU patients (adult surgical neonatal)

Widespread use of fluconazole emergence of less susceptible species

Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results

bull Dignosis based on clinical radiological microbiological and histopathological findings

bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Investigation included

ndash Microscopy

ndash Culture

ndash Serological test

ndash D-arabinitol detection

ndash Molecular assays

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids

bull C glabrata produces only yeast cells and

C albicans produces true hyphae in tissues

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection

bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites

bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication

bull Different susceptibility among Candida sp completed identification (speciation) before treatment

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis

bull Isolation of Candida sp from urine may indicate a serious infection

bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis

bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Limited usefulness

bull False positive (mucosal colonization superficial infection)

bull False negative (immunocompromised)

bull Antigen detection mannan insensitive

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

DETECTION OF D-ARABINITOL

bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)

bull It can be detected in serum or urine

bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR ASSAY

bull PCR detection of Candida DNA in blood serum LCS other specimens

bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid

bull False positive

bull Standardized commercial assays were not available

bull Can not be recommended in diagnosis of invasive candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

bull The Glucatell serum BG detection assay is highly sensitive and specific as a diagnostic adjunct for IFI

Zekaver Odabasi et al

Clinical Infectious Diseases 2004 39199ndash205

LABORATORY DIAGNOSTIC OF IFI

INVASIVE CANDIDOSIS

SPECIES DISTRIBUTION

bull The incidence of Candidosis infection in hospitalized patients in the USA has increased (1980-1989) 8 responsible for all bloodstream infections

bull In the 1990s incidence of Candida bloodstream infections decreased due to a decline of C albicans infections however the incidence rate of C glabrata increased

bull 1992-1993 1998-2000 overall incidence of Candida bloodstream infections increased

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

bull Invasive candidosis

ndash endogenous

ndash transmission from person to person (direct patient-patient transmission healthcare workers medical devices contaminated intravenous infusions)

bull Cross infection in ICU

ndash C albicans via hand of healthcare workers

ndash C parapsilosis via medical devices

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

bull Invasive candidosis

ndash Neutropenic cancer patients

ndash Stem cellorgan transplants recipients

ndash ICU patients (adult surgical neonatal)

Widespread use of fluconazole emergence of less susceptible species

Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results

bull Dignosis based on clinical radiological microbiological and histopathological findings

bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Investigation included

ndash Microscopy

ndash Culture

ndash Serological test

ndash D-arabinitol detection

ndash Molecular assays

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids

bull C glabrata produces only yeast cells and

C albicans produces true hyphae in tissues

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection

bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites

bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication

bull Different susceptibility among Candida sp completed identification (speciation) before treatment

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis

bull Isolation of Candida sp from urine may indicate a serious infection

bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis

bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Limited usefulness

bull False positive (mucosal colonization superficial infection)

bull False negative (immunocompromised)

bull Antigen detection mannan insensitive

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

DETECTION OF D-ARABINITOL

bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)

bull It can be detected in serum or urine

bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR ASSAY

bull PCR detection of Candida DNA in blood serum LCS other specimens

bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid

bull False positive

bull Standardized commercial assays were not available

bull Can not be recommended in diagnosis of invasive candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

INVASIVE CANDIDOSIS

SPECIES DISTRIBUTION

bull The incidence of Candidosis infection in hospitalized patients in the USA has increased (1980-1989) 8 responsible for all bloodstream infections

bull In the 1990s incidence of Candida bloodstream infections decreased due to a decline of C albicans infections however the incidence rate of C glabrata increased

bull 1992-1993 1998-2000 overall incidence of Candida bloodstream infections increased

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

bull Invasive candidosis

ndash endogenous

ndash transmission from person to person (direct patient-patient transmission healthcare workers medical devices contaminated intravenous infusions)

bull Cross infection in ICU

ndash C albicans via hand of healthcare workers

ndash C parapsilosis via medical devices

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

bull Invasive candidosis

ndash Neutropenic cancer patients

ndash Stem cellorgan transplants recipients

ndash ICU patients (adult surgical neonatal)

Widespread use of fluconazole emergence of less susceptible species

Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results

bull Dignosis based on clinical radiological microbiological and histopathological findings

bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Investigation included

ndash Microscopy

ndash Culture

ndash Serological test

ndash D-arabinitol detection

ndash Molecular assays

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids

bull C glabrata produces only yeast cells and

C albicans produces true hyphae in tissues

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection

bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites

bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication

bull Different susceptibility among Candida sp completed identification (speciation) before treatment

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis

bull Isolation of Candida sp from urine may indicate a serious infection

bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis

bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Limited usefulness

bull False positive (mucosal colonization superficial infection)

bull False negative (immunocompromised)

bull Antigen detection mannan insensitive

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

DETECTION OF D-ARABINITOL

bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)

bull It can be detected in serum or urine

bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR ASSAY

bull PCR detection of Candida DNA in blood serum LCS other specimens

bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid

bull False positive

bull Standardized commercial assays were not available

bull Can not be recommended in diagnosis of invasive candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

SPECIES DISTRIBUTION

bull The incidence of Candidosis infection in hospitalized patients in the USA has increased (1980-1989) 8 responsible for all bloodstream infections

bull In the 1990s incidence of Candida bloodstream infections decreased due to a decline of C albicans infections however the incidence rate of C glabrata increased

bull 1992-1993 1998-2000 overall incidence of Candida bloodstream infections increased

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

bull Invasive candidosis

ndash endogenous

ndash transmission from person to person (direct patient-patient transmission healthcare workers medical devices contaminated intravenous infusions)

bull Cross infection in ICU

ndash C albicans via hand of healthcare workers

ndash C parapsilosis via medical devices

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

bull Invasive candidosis

ndash Neutropenic cancer patients

ndash Stem cellorgan transplants recipients

ndash ICU patients (adult surgical neonatal)

Widespread use of fluconazole emergence of less susceptible species

Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results

bull Dignosis based on clinical radiological microbiological and histopathological findings

bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Investigation included

ndash Microscopy

ndash Culture

ndash Serological test

ndash D-arabinitol detection

ndash Molecular assays

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids

bull C glabrata produces only yeast cells and

C albicans produces true hyphae in tissues

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection

bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites

bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication

bull Different susceptibility among Candida sp completed identification (speciation) before treatment

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis

bull Isolation of Candida sp from urine may indicate a serious infection

bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis

bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Limited usefulness

bull False positive (mucosal colonization superficial infection)

bull False negative (immunocompromised)

bull Antigen detection mannan insensitive

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

DETECTION OF D-ARABINITOL

bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)

bull It can be detected in serum or urine

bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR ASSAY

bull PCR detection of Candida DNA in blood serum LCS other specimens

bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid

bull False positive

bull Standardized commercial assays were not available

bull Can not be recommended in diagnosis of invasive candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

bull The incidence of Candidosis infection in hospitalized patients in the USA has increased (1980-1989) 8 responsible for all bloodstream infections

bull In the 1990s incidence of Candida bloodstream infections decreased due to a decline of C albicans infections however the incidence rate of C glabrata increased

bull 1992-1993 1998-2000 overall incidence of Candida bloodstream infections increased

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

bull Invasive candidosis

ndash endogenous

ndash transmission from person to person (direct patient-patient transmission healthcare workers medical devices contaminated intravenous infusions)

bull Cross infection in ICU

ndash C albicans via hand of healthcare workers

ndash C parapsilosis via medical devices

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

bull Invasive candidosis

ndash Neutropenic cancer patients

ndash Stem cellorgan transplants recipients

ndash ICU patients (adult surgical neonatal)

Widespread use of fluconazole emergence of less susceptible species

Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results

bull Dignosis based on clinical radiological microbiological and histopathological findings

bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Investigation included

ndash Microscopy

ndash Culture

ndash Serological test

ndash D-arabinitol detection

ndash Molecular assays

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids

bull C glabrata produces only yeast cells and

C albicans produces true hyphae in tissues

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection

bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites

bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication

bull Different susceptibility among Candida sp completed identification (speciation) before treatment

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis

bull Isolation of Candida sp from urine may indicate a serious infection

bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis

bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Limited usefulness

bull False positive (mucosal colonization superficial infection)

bull False negative (immunocompromised)

bull Antigen detection mannan insensitive

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

DETECTION OF D-ARABINITOL

bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)

bull It can be detected in serum or urine

bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR ASSAY

bull PCR detection of Candida DNA in blood serum LCS other specimens

bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid

bull False positive

bull Standardized commercial assays were not available

bull Can not be recommended in diagnosis of invasive candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

bull Invasive candidosis

ndash endogenous

ndash transmission from person to person (direct patient-patient transmission healthcare workers medical devices contaminated intravenous infusions)

bull Cross infection in ICU

ndash C albicans via hand of healthcare workers

ndash C parapsilosis via medical devices

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

bull Invasive candidosis

ndash Neutropenic cancer patients

ndash Stem cellorgan transplants recipients

ndash ICU patients (adult surgical neonatal)

Widespread use of fluconazole emergence of less susceptible species

Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results

bull Dignosis based on clinical radiological microbiological and histopathological findings

bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Investigation included

ndash Microscopy

ndash Culture

ndash Serological test

ndash D-arabinitol detection

ndash Molecular assays

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids

bull C glabrata produces only yeast cells and

C albicans produces true hyphae in tissues

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection

bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites

bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication

bull Different susceptibility among Candida sp completed identification (speciation) before treatment

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis

bull Isolation of Candida sp from urine may indicate a serious infection

bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis

bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Limited usefulness

bull False positive (mucosal colonization superficial infection)

bull False negative (immunocompromised)

bull Antigen detection mannan insensitive

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

DETECTION OF D-ARABINITOL

bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)

bull It can be detected in serum or urine

bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR ASSAY

bull PCR detection of Candida DNA in blood serum LCS other specimens

bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid

bull False positive

bull Standardized commercial assays were not available

bull Can not be recommended in diagnosis of invasive candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

bull Invasive candidosis

ndash Neutropenic cancer patients

ndash Stem cellorgan transplants recipients

ndash ICU patients (adult surgical neonatal)

Widespread use of fluconazole emergence of less susceptible species

Note Intrinsic resistance (C glabrata and C krusei) to fluconazole factor in their emergence as pathogens

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results

bull Dignosis based on clinical radiological microbiological and histopathological findings

bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Investigation included

ndash Microscopy

ndash Culture

ndash Serological test

ndash D-arabinitol detection

ndash Molecular assays

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids

bull C glabrata produces only yeast cells and

C albicans produces true hyphae in tissues

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection

bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites

bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication

bull Different susceptibility among Candida sp completed identification (speciation) before treatment

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis

bull Isolation of Candida sp from urine may indicate a serious infection

bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis

bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Limited usefulness

bull False positive (mucosal colonization superficial infection)

bull False negative (immunocompromised)

bull Antigen detection mannan insensitive

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

DETECTION OF D-ARABINITOL

bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)

bull It can be detected in serum or urine

bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR ASSAY

bull PCR detection of Candida DNA in blood serum LCS other specimens

bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid

bull False positive

bull Standardized commercial assays were not available

bull Can not be recommended in diagnosis of invasive candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

LABORATORY DIAGNOSTIC

bull Diagnosis of invasive candidosis is difficult due to a variation and non specific clinical presentation difficulties in intrepreting laboratory examination results

bull Dignosis based on clinical radiological microbiological and histopathological findings

bull Microbiology positive blood culture budding yeast cellspseudohyphae in specimens obtained from normal sterile or abnormal sites (biopsy aspiration)

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

LABORATORY DIAGNOSTIC

bull Investigation included

ndash Microscopy

ndash Culture

ndash Serological test

ndash D-arabinitol detection

ndash Molecular assays

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids

bull C glabrata produces only yeast cells and

C albicans produces true hyphae in tissues

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection

bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites

bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication

bull Different susceptibility among Candida sp completed identification (speciation) before treatment

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis

bull Isolation of Candida sp from urine may indicate a serious infection

bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis

bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Limited usefulness

bull False positive (mucosal colonization superficial infection)

bull False negative (immunocompromised)

bull Antigen detection mannan insensitive

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

DETECTION OF D-ARABINITOL

bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)

bull It can be detected in serum or urine

bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR ASSAY

bull PCR detection of Candida DNA in blood serum LCS other specimens

bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid

bull False positive

bull Standardized commercial assays were not available

bull Can not be recommended in diagnosis of invasive candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

LABORATORY DIAGNOSTIC

bull Investigation included

ndash Microscopy

ndash Culture

ndash Serological test

ndash D-arabinitol detection

ndash Molecular assays

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids

bull C glabrata produces only yeast cells and

C albicans produces true hyphae in tissues

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection

bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites

bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication

bull Different susceptibility among Candida sp completed identification (speciation) before treatment

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis

bull Isolation of Candida sp from urine may indicate a serious infection

bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis

bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Limited usefulness

bull False positive (mucosal colonization superficial infection)

bull False negative (immunocompromised)

bull Antigen detection mannan insensitive

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

DETECTION OF D-ARABINITOL

bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)

bull It can be detected in serum or urine

bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR ASSAY

bull PCR detection of Candida DNA in blood serum LCS other specimens

bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid

bull False positive

bull Standardized commercial assays were not available

bull Can not be recommended in diagnosis of invasive candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

MICROSCOPY

bull Typical budding yeast cells pseudohypahetrue hyphae of Candida species found in tissue biopsy normal sterile body fluids

bull C glabrata produces only yeast cells and

C albicans produces true hyphae in tissues

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection

bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites

bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication

bull Different susceptibility among Candida sp completed identification (speciation) before treatment

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis

bull Isolation of Candida sp from urine may indicate a serious infection

bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis

bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Limited usefulness

bull False positive (mucosal colonization superficial infection)

bull False negative (immunocompromised)

bull Antigen detection mannan insensitive

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

DETECTION OF D-ARABINITOL

bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)

bull It can be detected in serum or urine

bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR ASSAY

bull PCR detection of Candida DNA in blood serum LCS other specimens

bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid

bull False positive

bull Standardized commercial assays were not available

bull Can not be recommended in diagnosis of invasive candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

CULTURE

bull Isolation of Candida sp from sputum and faecal specimens can not be considered diagnostic of infection

bull Deep-seated infection Candida sp isolated from blood or normally sterile body fluids tissuesaspirates from other closed sites

bull Specimens should be examined as soon as possible after collection to avoid missinterpretation due to other organisms multiplication

bull Different susceptibility among Candida sp completed identification (speciation) before treatment

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

CULTURE

bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis

bull Isolation of Candida sp from urine may indicate a serious infection

bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis

bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Limited usefulness

bull False positive (mucosal colonization superficial infection)

bull False negative (immunocompromised)

bull Antigen detection mannan insensitive

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

DETECTION OF D-ARABINITOL

bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)

bull It can be detected in serum or urine

bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR ASSAY

bull PCR detection of Candida DNA in blood serum LCS other specimens

bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid

bull False positive

bull Standardized commercial assays were not available

bull Can not be recommended in diagnosis of invasive candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

CULTURE

bull Blood cultures are positive in no more than 50 of neutropenic patients with dissemintaed candidosis or 80 of patients with endocarditis

bull Isolation of Candida sp from urine may indicate a serious infection

bull Isolation of Candida sp from LCS evidence for diagnosis of meningitis

bull Candida sp isolated from BAL in a patient with pulmonary infiltrates not sufficient for diagnosing candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Limited usefulness

bull False positive (mucosal colonization superficial infection)

bull False negative (immunocompromised)

bull Antigen detection mannan insensitive

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

DETECTION OF D-ARABINITOL

bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)

bull It can be detected in serum or urine

bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR ASSAY

bull PCR detection of Candida DNA in blood serum LCS other specimens

bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid

bull False positive

bull Standardized commercial assays were not available

bull Can not be recommended in diagnosis of invasive candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

SEROLOGICAL TESTS

bull Limited usefulness

bull False positive (mucosal colonization superficial infection)

bull False negative (immunocompromised)

bull Antigen detection mannan insensitive

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

DETECTION OF D-ARABINITOL

bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)

bull It can be detected in serum or urine

bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR ASSAY

bull PCR detection of Candida DNA in blood serum LCS other specimens

bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid

bull False positive

bull Standardized commercial assays were not available

bull Can not be recommended in diagnosis of invasive candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

DETECTION OF D-ARABINITOL

bull D-arabinitol is produced by most of Candida sp (except C krusei and may be C glabrata)

bull It can be detected in serum or urine

bull Candidemia increased of serum D-arabiniitol-creatinine ratios has still to achieve widespread clinical use

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR ASSAY

bull PCR detection of Candida DNA in blood serum LCS other specimens

bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid

bull False positive

bull Standardized commercial assays were not available

bull Can not be recommended in diagnosis of invasive candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

MOLECULAR ASSAY

bull PCR detection of Candida DNA in blood serum LCS other specimens

bull Promising tools for rapid diagnosis of Candida sp from tissue or body-fluid

bull False positive

bull Standardized commercial assays were not available

bull Can not be recommended in diagnosis of invasive candidosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

The sources of yeast isolated

bull Sputum 49

bull Urine 13

bull Vaginal swab 9

bull Blood 8

bull Bronchial lavage 7

2010

bull Sputum 62

bull Vaginal swab 16

bull Blood 14

bull Urine 9

bull Bronchial lavage 5

2011

bull Sputum 102

bull Urine 18

bull Blood16 bull Bronchial lavage 12 bull Vaginal swab 11

2012

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

Candida spp isolated from blood

2010 2011 2012

C albicans 1 1 3

C parapsilosis 3 4 1

C pelliculosis 1 0 0

C lusitaniae 1 0 0

C tropicalis 0 2 10

C glabrata 0 1 0

C gulliermondii 0 1 1

Candida spp 1 0 0

7 9 15

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

Resistance pattern of Candida spp isolated from blood

2010 2011 2012

FCA ITR VRC FCA ITR VRC FCA ITR VRC

C albicans 17 115 115 115

C glabrata 19

C tropicalis 215 315 115

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

INVASIVE ASPERGILLOSIS

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

bull Incidence varies in ICU 58

bull In ICU risk factors including chronic obstructive pulmonary diseases and liver failure

bull Positive respiratory culture uncertain

bull Low sensitivity and specificity

bull Detection of galactomannan in BAL promising results

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

LABORATORY DIAGNOSTIC

bull Microscopy

bull Culture

bull Serological tests

bull Molecular diagnostics

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

MICROSCOPY

bull Histopathological examination of tissue biopsies immunohistochemical staining procedure

bull Non-pigmented septate hyphae dichotomous branching

bull Direct examination of sputum may help in diagnosing invasive aspergillosis BAL is better

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

A fumigatus

Richard C Barton Hindawi Publishing Corporation Scienti1048853 ca Volume 2013 Article ID 459405 29 pages httpdxdoiorg1011552013459405

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

CULTURE

bull Not sensitive interpreted with caution (contamination)

bull Recovered from blood LCS

bull Aspergillus isolated from BALsputum obtained from patients with a pulmonary infiltrates indicative of infection

bull A fumigatus can be isolated from sputum of patients suffer from allergic bronchopulmonary aspergillosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

SEROLOGICAL TESTS

bull Antobodies againts Aspergillus often helpful in the diagnosis of aspergillosis in non immunocompromised patients

bull Immunodiffusion indirect haemagglutination ELISA

bull The role is still uncertain

bull Positive ID test is not a proof of infection negative result can not exclude the diagnosis

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

SEROLOGICAL TESTS

bull Antigen detection (galactomannan) in blood and other body fluids rapid diagnosis of aspergilllosis in immunocompromised patients

bull Galactomannan fluctuate during the course of the infection useful if performed in a regular basis (twice per week)

bull Negative not exclude the diagnosis

bull ELISA for the detection of galactomannan has been approved for diagnostic use (the accuracy was moderate)

Christopher D Pfeiffer et al 2006

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

MOLECULAR DIAGNOSTICS

bull PCR for detection of Aspergillus in blood BAL serum etc

bull Appear promising for rapid detection of Aspergillus directly on tissue or body fluids but

ndash False positive

ndash No standardized commercial method

bull Routine use of PCR can not be recommended

Richardson MD and Warnock DW 2003 Fungal infection diagnosis and management

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

Mark Reinwald et al

bull ldquoTreatment with mould-active antifungals prior to BAL sampling significantly decreases the performance of the Aspergillus PCR assay in haematological patients if BAL was performed after administration of more than one antifungal agentrdquo

bull ldquoThe effect of antifungal therapy on other surrogate parameters suggests reduced sensitivity rates with antifungal therapy for GM whereas no influence of antifungal therapy on β-D-glucan performance in a (single-centre retrospective) study was describedrdquo

J Antimicrob Chemother 2012 67 2260ndash2267

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI

CONCLUSION

bull Various detection methods of agents responsible for invasive fungal infections have been developed

bull The lack of sensitivity and specificity of the test uncertainty in diagnosing IFI decrease successful treatment

bull Need further studies to improve and develop laboratory diagnostic tool for IFI