seminar IDI Barabai 9 Mei 2015 revisi (dr. Yanti Sp.PK).pptx

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LABORATORY FINDING IN DENGUE VIRUS

INFECTION

dr. Faizah Yunianti Sp.PKRSUD. H.Damanhuri - Mitra Laboratorium

Barabai

Struktur virus dengue

Virus dengue

DEN 3

DEN 2 DEN 4

2

DEN 1

Famili Flavivirida

e

ArbovirusØ 30 nm

2

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penyakit flu, demam tifoid, demam chikungunya, leptospirosis, malaria

Tidak khas

Sulit diagnosisTerutama pada masa

awal demam (3-4 hari)

Manifestasi klinis

Beragam : asimtomatik, undifferentiated febrile illness, demam dengue, demam berdarah dengue (DBD) ,perembesan plasmasyok hipovolemik (SSD),kematian

Infeksi virus dengue

Manifestasi laboratoris

Trombositopenia ringan sampai dengan berat,

berbagai kombinasi kehadiran NS1, IgG dan IgM dan variasi kadar sitokin.

4

Peran pemeriksaa

n laboratoriu

m

4

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Detection of the virus / virus isolation (culture)PCRDetection of antidengue antibody or antigen

(serology)LeukopeniaLymphocytosisThrombocytopeniaRaised of hematokrit

Laboratory Diagnosis

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Leukopenia,Thrombocytopenia, Raised of hematokrit

lymphocytosis

Puskesmassederhana

bilik hitung, reangensia mikroskop

Rumah Sakit DL Otomatikbilik hitung, reangensia mikroskop

Hapusan darah

True/ False

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Nama : kasus 1 Umur : YPARAMETER HASIL NILAI NORMAL

Hemoglobin (Hb) 12,1 12,0 – 17,4g/dL

Eritrosit (RBC) 3,9 4,0 - 5,5 X 1012 / L

Hematokrit (Ht) 49,0 36,0 – 52,0 %

MCV 81 76 - 96 fL

MVH 29,0 27,0 – 32,0 pg

MCHC 33,0 31,0 – 38,0 g/dL

RDW CV 14,3, 0,0 – 16,0%

Lekosit (WBC) 3,3 5,0 – 10,0 X 109/ L

Eosinofil 1,0 0,0 – 2,0 %

Basofil 0,0 0,0 – 1,0%

Neutrofil 34 50 – 70 %

Limfosit 65 20 - 40%

Monosit 6 2 – 8 %

Trombosit (Plt) 57 150 – 450 X 109/ L

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Eritrosit (RBC) 3,9 4,0 - 5,5 X 1012 / L

Hematokrit (Ht) 36 36,0 – 52,0 %

MCV 81 76 - 96 fL

MVH 29,0 27,0 – 32,0 pg

MCHC 33,0 31,0 – 38,0 g/dL

RDW CV 14,3, 0,0 – 16,0%

Lekosit (WBC) 3,3 5,0 – 10,0 X 109/ L

Eosinofil 1,0 0,0 – 2,0 %

Basofil 0,0 0,0 – 1,0%



Monosit 6 2 – 8 %

Trombosit (Plt) 57 150 – 450 X 109/ L

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Eritrosit (RBC) 3,9 4,0 - 5,5 X 1012 / L

Hematokrit (Ht) 36 36,0 – 52,0 %

MCV 54 76 - 96 fL

MVH 29,0 27,0 – 32,0 pg

MCHC 33,0 31,0 – 38,0 g/dL

RDW CV 14,3, 0,0 – 16,0%

Lekosit (WBC) 4.7 5,0 – 10,0 X 109/ L

Eosinofil 1,0 0,0 – 2,0 %

Basofil 0,0 0,0 – 1,0%



Monosit 6 2 – 8 %

Trombosit (Plt) 165 150 – 450 X 109/ L

Virus IsolationDefinitive test for dengue infectionLab equipped with tissue culture facilitiesUseful only at early phase of illness , blood

collected before day 5 of illness ( before the formation of Neutralizing antibodies)

During febrile illnessVirus can be isolated from serum, plasma and

leucocytesPost mortem specimens

Expensive and time consuming (2 weeks to complete

Polymerase Chain Reaction (PCR)

Diagnosis of dengue infection in the early phase (< 5 days of illness)

Sensitivity 100% in the first 5 days of illness, reduced to 70% by day 6

Determine dengue serotype

Polymerase Chain Reaction (PCR)Limitation

Available in certain facilities and trained personal

Test is expensiveSpecimens requires special storage

temperatures and short transportation , time between collection and extraction

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Deteksi AntigenNS1 (Non struktural Antigen 1)

Deteksi AntibodiIg M dan Ig G

Serologi

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Mencari bagian tertentu dari virus Dengue yang dapat menimbulkan penyakit baik berupa peptida ataupun asam nukleat

Deteksi Antigen

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tidak menunjukkan perbedaan pada infeksi primer maupun sekunder, dan beberapa kasus masih (+) pada fase konvalesen

marker deteksi dini infeksi dengue primer-sekunder

protein non-struktural yang disekresikan di permukaan sel terinfeksi

ditemukan dalam sirkulasi darah selama fase akut infeksi virus dengue

Antigen NS1

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glikoprotein 46-50 kDa terdapat di permukaan sel (membrane-associated /mNS1 ) & secreted form (sNS1)bentuk homodimer, partially hydrophobic nature bukan bagian dari struktur virussel yang terinfeksi di ekspresikan pada permukaan 1 dari 7 protein NS yang dihasilkan selama replikasi virusprotein yang disekresikan melalui cellular secretory pathway ke permukaan sel

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antigen NS1 dengue

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Dengue IgM Antibody IgM titre is higher in primary compared to

secondary infectionPeak at about 2 weeks and undetectable level

by 60 – 90 days. Recommended if dengue IgM is still

negative after day 7 in secondary dengue

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Dengue IgM Antibody Primary Dengue

55% detectable between 4-7 days100% after day 7

Secondary 78% after day 728% of secondary dengue infections were

undiagnosed when IgM was the only test performed

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Dengue IgG Antibody Primary and secondary dengue

IgG was detected in 100% after day 7 of illness

If Ig G was detected before day 7, that is a secondary infection

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Ig G Primary

WHO, 2009

Ig G Secondary

Ig M Secondary

Ig M Primary

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Interpretasi Hasil Ig m dan Ig GNo.Urut

Hasil InterpretasiIgG IgM

1. positif positif Dengue sekunder2. negatif positif Dengue primer3. positif negatif Dugaan dengue

sekunder

4. negatif Negatif Non-Dengue/ Primer amat awal. Retest setelah 4-7 hari

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Dengue Rapid testFalse positiveCross react with

Flavivirus-Non-flavivirus – malaria, leptospirosis,

toxoplasmosis, syphilisConnective Tissue Disorder

False negative low titre antigen/antibody

* ELISA METHODFOR DETERMINE

TITRE OF ANTIGEN/ANTIBOD

Y

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Keuntungan pemeriksaan antigen NS1deteksi infeksi dengue pada fase awaldemam (hari ke-1 sampai ke-5 sakit)tanpa menunggu terbentuknya antibodiSensitivitas pemeriksaan NS1 (88,7%) lebih tinggi

dibandingkan kultur virus (68,1%) dan PCR (66,7%).

Pemeriksaan IgG dan IgM antidenguetetap diperlukan untuk membedakan infeksi primer

atau infeksi sekunder

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KESIMPULAN

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ReferensiHandbook of Clinical Managemen of Dengue . WHO. 2012Nilai Novel Kombinasi Nilai Antingen NS1, Antibodi IgM dan IgG

Anti Dengue Dalam Memprediksi Outcome Infeksi Virus Dengue di Makassar. Rachmat Latief. 2010

Diagnosis cepat demam berdarah dengue. Danny wiradharma. 1999

Pemeriksaan Laboratorium Pada Demam Berdarah Dengue. Aryati. 2009

Laboratory Investigation. Salmah IdrisSerology Diagnosis of Dengue Infections. ECDC training

Workshop on laboratory diagnosis of dengue virus infectionsBerlin, 23‐27 January 2012

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TERIMA KASIH

Documents

seminar IDI Barabai 9 Mei 2015 revisi (dr. Yanti Sp.PK).pptx