第1页：Experiment five Isolation and
Experiment five Isolation and
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第2页：Isolation and Identification
Isolation and Identification
- Specimen
- Isolation and identification of virus
- Serological diagnosis of viral infection
- Rapid diagnosis of virus infections
第3页：Isolation and Identification
Isolation and Identification
- Isolation and identification of virus
- Isolation of virusEmbryonatedEggsLaboratoryAnimalsTissueCulture
- Quantitation of virus50%tissuecultureinfectiousdose(TCID50)Plaque forming assay
第4页：Isolation and Identification
Isolation and Identification
- Serological diagnosis of viral infection
- Neutralization test(NT)
- Complement fixation test(CF)
- Titration of complement
第5页：Isolation and Identification
Isolation and Identification
- Rapid diagnosis of virus infections
- Identification virus particle
- Detection of viral antigens
- Detection of viral nucleic acid
- Diagnosis of virus infections bydetecting IgM antibodies
- Other methods
第6页：Isolation and Identification
Isolation and Identification
- Purpose：
- To obtain information about microbiologicaldiagnosis methods of influenza virus
- Procedure
- Acquiring and managing specimen
- Isolation and Identification of influenza virus
- serological diagnosis of influenza virus
第7页：Isolation and Identification
Isolation and Identification
- Acquiring and managing specimen
- Throat washing or gargling are obtained within 3days after infection
- Test at once or stored frozen.
- Centrifuge the washings 15 min at 3000 rpm,Harvest 1 ml of supernatant.
- Add antibiotic to prevent bacterial contamination.
第8页：Hemagglutination inhibition
Hemagglutination inhibition
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第9页：Isolation and Identification
Isolation and Identification
- Isolation of influenza virusMATERIALS
- 1.Throat washings or garglings
- 2.Embryos (within 9 to 10-day-old)
- 3.0.5% chicken RBC suspension, physiologicalsaline
- 4. Scissors, forceps, drill, syringe, tubes, egg-checking box, 75% ethanol, tincture of iodine
第10页：Markthelocationofembryoairsacunderegg-checkingbox,sterilizetheshellwithtinctureand75%ethanol,drillasmallholenearthemarkedshell,inoculate0.2mlofvirusinoculumintotheallantoiccavitybypushingtheneedlejustbelowthesurfaceoftheshell
Markthelocationofembryoairsacunderegg-checkingbox,sterilizetheshellwithtinctureand75%ethanol,drillasmallholenearthemarkedshell,inoculate0.2mlofvirusinoculumintotheallantoiccavitybypushingtheneedlejustbelowthesurfaceoftheshell
- Markthelocationofembryoairsacunderegg-checkingbox,sterilizetheshellwithtinctureand75%ethanol,drillasmallholenearthemarkedshell,inoculate0.2mlofvirusinoculumintotheallantoiccavitybypushingtheneedlejustbelowthesurfaceoftheshell,
第11页：Hemagglutination test, HT
Hemagglutination test, HT
- MATERIALS
- Virus:theallantoicfluidthatcontainsinfluenzavirus;
- physiologicalsaline;
- 0.5%chickenRBCsuspension.
- 20-wellplasticdisposablemicrotiterplatesandglasspipettes.
第12页：Materialswell No.(ml)1 2 3 4 5
Materialswell No.(ml)1 2 3 4 5
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第13页：Hemagglutination test, HT
Hemagglutination test, HT
第14页：Hemagglutination test
Hemagglutination test
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第15页：Hemagglutination test (HT)
Hemagglutination test (HT)
- RESULTRecordthehemagglutinationasthefollowings:++++:AlltheRBChadbeenagglutinated.+++:75%ofRBChadbeenagglutinated++:50%ofRBChadbeenagglutinated.+:25%ofRBChadbeenagglutinated.-:NoRBCwasagglutinated.
第16页：Hemagglutination inhibition
Hemagglutination inhibition
- MATERIALS
- Patient‘s serum: deactivated 30 min at 56C;
- influenza virus: 4 HA units;
- 0.5% chicken RBC suspension;
- physiological saline
- 20-well microtiter plates, pipettes and tubes.
第17页：Materialswell No.(ml)1 2 3 4 5
Materialswell No.(ml)1 2 3 4 5
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本页原文为图示化表格，文字提取后字段粘连，未能安全切分为结构化表格，建议对照课件原图人工核对后补充。
第18页：
第19页：Hemagglutination inhibition
Hemagglutination inhibition
- RESULTAssesshemagglutinationasabove,thehemagglutinationinhibition(HI)titeristhereciprocalofthehighestdilutionofthepatient'sserumwhichshowscompleteinhibitionofagglutination.1:40dilutioninTableistheHItiter.
第20页：
第21页：Isolation and Identification
Isolation and Identification
- Identification of influenza virus
- Hemagglutination inhibition test (HI)---subtype
- Complement fixation test (CF)---type
