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Virus neutralization test


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Virus neutralization test

  2. 2. VIRUS NEUTRALIZATION TEST Introduction  Serological method that detects the presence of viral neutralizing antibodies.  The antibodies bind to the viral particles • Monoclonal and polyclonal antibodies  Block viral infectivity-No cell infection SHAREEFNGUNGUNI6/1/2017 2
  3. 3. VNT cont..,  VNT are conducted by :  mixing dilutions of antibodies with standardized amount of virus, incubating them and cultured into cells, eggs or animals to have a clear cytopathic effect observation  CPE effects:  Rounding of cells  Change in texture-granular/hyaline/glassy  Formation of syncytium SHAREEFNGUNGUNI6/1/2017 3
  4. 4. VNT cont..,  STEPS • First step: • Serial dilutions of serum(virus-neutralizing Ab?)+ known virus • Incubated for 1-2hr at 37 degrees Celsius • Second step; • Cell culture inoculated with mixture • Seal the plates & Incubated at 37 degrees Celsius • Direct microscope SHAREEFNGUNGUNI6/1/2017 4
  5. 5. VNT cont., • Illustrations using diagrams Copyright 2009, John Wiley & Sons, Inc. Copyright 2009, John Wiley & Sons, Inc. Copyright 2009, John Wiley & Sons, Inc.. right 2009, John Wiley & Sons, Inc. 6/1/2017 5 SHAREEFNGUNGUNI
  6. 6. VNT cont.., • Expectations • No Ab(in serum)+virus no neutralization CPE • Ab(in serum)+virus neutralization No CPE • Cells remain intact SHAREEFNGUNGUNI6/1/2017 6
  7. 7. VNT cont.., -pro’s: • Can be specific • Sensitive • Vaccine production/immunological studies -con’s: • Very slow(days) • Intensive • Require skilled people • Depend on cell line-use of wrong cell lines may assume that the antibodies have neutralization ability when not, or may seem to be ineffective when they actually have the neutralization abilities. SHAREEFNGUNGUNI6/1/2017 7
  8. 8. Conclusion All in All, viral infections can be diagnosed in laboratory by many methods and the gold standard one is neutralization tests since it is a technique to which all other test methods have been compared with. Although the technique is highly sensitive and specific in detection of antibodies, it has disadvantages. SHAREEFNGUNGUNI6/1/2017 8
  9. 9. REFERENCES 1) Carter,J.,and Saunders,V.(2007).Virology:Principles and Applications.John Willey & Sons,Ltd.England,UK. 2) Flint,S.J.,Enquist,L.W.,Racaniello,V.R.,and Skalka,A.M.(2004).Principles of virology:Molecular Biology,Pathogenesis and Control of Animal Viruses.2nd Edition.ASM press,Washington DC,USA. 3) Murphy, F.A.,Gibss ,J.P.,Horzinek.M.C.,and Studdert,M.J.(1999).Veterinary Virology.2nd edition.Elsevier,Inc.Califonia,USA. SHAREEFNGUNGUNI6/1/2017 9