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Table Of Contents

A.Introduction
B.Goals of the proposed WHO Animal Influenza Network
C.Background information on surveillance for animal influenza
D.Transmission of influenza viruses from lower animals to humans
A.Collection of specimens
Introduction
Specimens for diagnosis
Transport and storage of specimens
Prepare sample vials
Preparing to take samples
Sera collection for influenza diagnosis and surveillance
Transporting specimens to the laboratory
B.Isolation of influenza viruses
Processing clinical material for virus isolation
C.Isolation in cell culture
Preparation of media and reagent formulas
Preparation of MDCK cells in tissue culture flasks
Inoculation of cell culture
D.Inoculation of embryonated chicken eggs
Procedure
Precautions
Identification of contamination
E.Identification of influenza isolates by hemagglutination inhibition
Hemagglutination and Hemagglutination Inhibition Test
Limitations
Procedure for serologic diagnosis
Materials required
Neuraminidase assay (NA assay)
Neuraminidase inhibition assay (NAI)
Screening of sera for evidence of NA antibodies
H.Neutralization assay for antibody detection
Procedure for influenza virus neutralization assay
Quality Control
Procedure of the Neutralisation Assay
RNA extraction from viral isolate
Synthesis of cDNA
PCR reaction
Agarose Gel Electrophoresis of the PCR products
Chemicals and enzymes used in the molecular assays
Method
Definitions
Safety
Materials
Test method
A.Forms
B.Definitions
D.Laboratory safety
E.Hazardous chemicals
F.Alternate protocols
Influenza HAI serum treatment
Influenza HAI standardization of RBCs
Alternative method: Micro-Neutralisation Assay
Background and surveillance
Collection of clinical specimens
Diagnosis of respiratory virus infections
Hemagglutination - hemagglutination inhibition
Neutralization assay
Molecular analysis
Detection of influenza virus Type A
Avian viruses
H5N1 viruses
H9N2 viruses
Influenza in pigs
Equine influenza
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WHO Manual Influenza 2002

WHO Manual Influenza 2002

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Published by: rwbbarton on Jun 23, 2011
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11/09/2012

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