Eye Infection

LABORATORY EXAMINATION IN THE ETIOLOGICAL DIAGNOSIS OF

EYE FOR INFECTION

WINARTODEPT. of OPHTHALMOLOGY

FAC. of MEDICINE, DIPONEGORO UNIVERSITY / DR KARIADI HOSPITAL

S E M A R A N G

1. Sites with normal flora : a. Function : - part of innate immunity - contribute to body normal function b. If disturb dysfunction causes a diseases

e.g.: skin (palpebra), conjunctivae, cornea, lacrimal system.

2. Steril sites :e.g. COA, COP, vitreous, subcutaneous tissue,

subconjungtival tissue

Indogenous Microflora of Human Eye

INTRODUCTION

Site with normal flora

TIDAK MEMPENGARUHI

TAJAM PENGLIHATAN

Hordeolum eksternum Hordeolum internum

Kalazion

• Kalazion• Jaringan granuloma

pada tarsus inferior

Selulitis (anterior) palpebra

• Palpebra bengkak• Tanda radang

pada palpebra

EMERGENCY

DAPAT MEMPENGARUHI

TAJAM PENGLIHATAN

Trikiasis

• Silia atas tumbuh ke arah dalam : kornea atau konjungtiva

• Kornea atau konjungtiva teriritasi

• Akibatnya terjadi Keratitis atau Konjungtivitis

Konjungtivitis bakteri

• Konjungtivitis bakteri – Sekret

mukopururulen– Konjungtiva kemotik– Injeksi konjungtiva

Keratitis marginal

• Abses berbentuk cincin di tepi kornea

• Jernih antara keratitis dan limbus

Pemakai lensa kontakKERATITIS

DECORATIVE / COSMETIC

Konjungitivitis purulenta

• Konjungtivitis purulenta gonorrhoe :– Konjungtiva

kemotik dan kasar– Sekret purulen

EMERGENCY

Konjungtivitis virus

• Konjungtivitis virus– Injeksi

konjungtival– Sekret sereous– Perdarahan

subkonjungtiva ( subakut )

Herpes zooster oftalmikus

• Herpes zoster oftalmikus

• Stadium penyembuhan

Keratitis dendritik

• Infiltrat dengan batas seperti cabang-cabang

• Disebabkan Herpes simpleks

Trakoma • Konjungtivitis

trakoma– Folikel pada

konjungtiva tarsal

– Panus• Infiltrat limbus

atas• Neovaskularisasi

di atas

Keratitis lagoftalmos

• Lagoftalmos pada penderita eksoftalmus goiter

• Keratitis di bagian bawah akibat mata tidak tertutup waktu tidur

Ulkus sentral

• Ulkus dengan neovaskularisasi dari limbus

EMERGENCY

Ulkus atau abses kornea + hipopion

• Kemotik + injeksi siliar

• Abses kornea• Hipopion di dalam

bilik mata depan

EMERGENCY

Endoftalmitis

• Injeksi siliar• Massa supuratif di

dalam bilik mata depan

EMERGENCY

Nebula kornea

• Kekeruhan tipis pada kornea

• Batas kabur• Tanda radang

negatif

Leukoma kornea

• Kekeruhan dengan - Batas tegas - Mata tenang

Infection ?

Inflammation ?

Clinical Diagnosis / Suspicion:

Infection

Clinical Diagnosis / Suspicion:

1. Bacteria2. Fungus3. Virus

Infection

Clinical Diagnosis / Suspicion:

1. Bacteria2. Fungus3. Virus

1. Isolation2. Susceptibility test3. Antigen detection4. Immunologic respons5. Molecular diagnosis

IMMUNOLOGIC RESPONS

HUMORAL

CELLULAR

IgG, IgM, IgA, IgE, IgDCytokines

DIAGNOSTICS

IMMUNOLOGIC (RESPONS) DETECTION

HUMORAL

IgG, IgM, IgA, IgE, IgD

DIAGNOSTICS

1. Paired sera: acute and convalescence

2. Rises of titer3. Endemic titer

INPUT OUTPUTPROCESS

GARBAGE IN GARBAGE OUT

MICROBIOLOGY PROCESSES

INPUT OUTPUTPROCESS

CLINICAL SPECIMEN

MICROBIOLOGY PROCESSES

INPUT OUTPUTPROCESS

OPHTHALMOLOGIST

SPECIMEN COLLECTION

MICROBIOLOGY PROCESSES

INPUT OUTPUTPROCESS

CLINICAL MICROBIOLOGIST

SPECIMEN PROCESSING & REPORTING

MICROBIOLOGY PROCESSES

INPUT OUTPUTPROCESS

OPHTHALMOLOGIST

CLINICAL MICROBIOLOGIST

SPECIMEN COLLECTION

SPECIMEN PROCESSING & REPORTING

MICROBIOLOGY PROCESSES

OPHTHALMOLOGIST

CLINICAL MICROBIOLOGIST

Request Form :

Microbiology lab. :

1. Administration

2. Specimen inspection

3. Lab. Examination

4. Report

Main objectives : to serve better to the patients

communication

Clinical Diagnosis

(Request form)

Sample

Staining

Isolation & identification

Susceptibility test

Preliminary report

Final report

Serology results

MICROBIOLOGY EXAMINATION

Serology detection

Morphology / microscopy

Species nameAntibiogram

IgM & IgG

Rapid & mol. diagnosis IF, PCR, RFLP, Blotting, etc

SPECIMEN COLLECTION

IS VERY IMPORTANT

PROPER SPECIMEN

AVOID CONTAMINATION

BEFORE ANTIBIOTICS TR/

Ulkus Kornea

X

LABORATORY EXAMINATION

Scrapping Staining: Gram, KOH, Calcuflor white

Culture & Sensitivity test

Fungus culture: SDA

Acanthamoeba culture: non nutrient medium

BA, Chocolate agar, Mc Conkey

TYPE OF SPECIMENS

SWAB: - ConjungtivitisSCRAPPING: - Conjungtivitis to study

epithelial cells (inclusion bodies) - Corneal ulcers: - fungus - viral (HSV1) - acanthamoebaPUS: hordeolum SKIN SCRAPPING’S: HSV1, mycoses

CORNEAL ULCER SPECIMEN

Local Analgesics:

- suppressed bacteria

- get more specimens

Limited specimens:

- culture : direct inoculation

enrichment broth isolation

- staining

STAINING

1. GRAM

2. GIEMSA

Bacterial MorphologyGram (+) / (-) p.m.n. choose antibiotics

Inclusion bodiesCellular respons (pmn/lymphocyte) Viral / chlamydial infection Acute / chronicAllergy

3. KOH Filament, yeast, spores

4. Imm-Fluoresens HSV

Corneal scraping (Masson trichrome) cyst of Acanthamoeba (arrow). Dendritiform epithelial lesions of patient with Acanthamoeba keratitis.

ISOLATION & SENSITIVITY TEST

CULTURE

Minimal using 2 type of media: - non inhibitory media- inhibitory media

Isolated colony

IdentificationColony morphologyStainingBiochemical reaction, etc

Primary plating = isolation

Identification = Biochemistry reaction

Antibiotic Sensitivity Test

SensitiveIntermediate sensitiveResistant

MIC : minimum inhibitory concentration

MBC : minimum bacteri- cidal concentration

Disc diffusion method :

Dilution method :

Causative organismConjungtivitis :

S pneumoniae, S aureusH influenzaeC trachomatis (inclusion & trachoma)Others : neisseriaViral : adenovirus.

Keratitis : S pneumoniae, S aureus, S pyogenes

Pseudomonas, Enterobacteriaceae, othersAcanthamubaViral : HSV, Herpes zoozter.

Endophthalmitis :S aureus, Pseudomonas, S pneumoniaeP acnesN meningititidsOthers :

Periorbital sellulitis : S aureus, S pyogenes, S pneumoniae,

H influenzaeClostridium

WHEN TAKING SPECIMEN FOR CULTURE ???

x

x

x

Immediate AB, culture if no respond

Immediate culture, AB wait culture

Immediate culture & AB

AB culture Bacteria die

Disease progress

Best choice