Processing of Clinical Specimen for Immunofluorescence staining & Viral

culture

Exercise II – Day 4 Virology Section

Department of MicrobiologyJIPMER

XIII NATIONAL WORKSHOP ON “DIAGNOSTIC METHODS IN CLINICAL MICROBIOLOGY”

8th – 11th October, 2014

XIII National Workshop on Diagnostic Methods in Clinical Microbiology, JIPMER

2

Introduction

Viral detection methods:•Staining

Direct stainingImmunofluorescence staining

• Culture Cell culture Animal / embryonated egg inoculation

• Enzyme immunoassays• PCR methods

XIII National Workshop on Diagnostic Methods in Clinical Microbiology, JIPMER

3

Objective

To observe and understand the steps in processing a clinical

specimen for immunofluorescence staining and viral culture

XIII National Workshop on Diagnostic Methods in Clinical Microbiology, JIPMER

4

Steps – Processing of specimen

Sample

receipt

Vortex &aliqu

ot

Centrifug

e

Use for Viral culture

Use for Immunofluorescence staining

XIII National Workshop on Diagnostic Methods in Clinical Microbiology, JIPMER

5

IFA

XIII National Workshop on Diagnostic Methods in Clinical Microbiology, JIPMER

6

Positive IFA- Adenovirus Negative IFA

XIII National Workshop on Diagnostic Methods in Clinical Microbiology, JIPMER

7

Detection of virus growth in cell cultures

Cytopathic effect

Metabolic inhibition

Hemadsorption

Interference

Transformation

Immunofluorescence

Normal cell line – Hep2 CPE- Adenovirus

XIII National Workshop on Diagnostic Methods in Clinical Microbiology, JIPMER

Grape like clusters of round cells

Polygonal cells

Thank you

XIII National Workshop on Diagnostic Methods in Clinical Microbiology, JIPMER