Reproducibility of serologic assays for influenza virus A (H5N1)

Emerg Infect Dis. 2009 Aug;15(8):1252-9. doi: 10.3201/eid1508.081754.

Abstract

Hemagglutination-inhibition (HI) and neutralization are used to evaluate vaccines against influenza virus A (H5N1); however, poor standardization leads to interlaboratory variation of results. A candidate antibody standard (07/150) was prepared from pooled plasma of persons given clade 1 A/Vietnam/1194/2004 vaccine. To test human and sheep antiserum, 15 laboratories used HI and neutralization and reassortant A/Vietnam/1194/2004, A/turkey/Turkey/1/2005 (clade 2.2), and A/Anhui/1/2005 (clade 2.3.4) viruses. Interlaboratory variation was observed for both assays, but when titers were expressed relative to 07/150, overall percentage geometric coefficient of variation for A/Vietnam/1194/2004 was reduced from 125% to 61% for HI and from 183% to 81% for neutralization. Lack of reduced variability to clade 2 antigens suggested the need for clade-specific standards. Sheep antiserum as a standard did not reliably reduce variability. The World Health Organization has established 07/150 as an international standard for antibody to clade 1 subtype H5 and has an assigned potency of 1,000 IU/ampoule.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Viral* / analysis
  • Antibodies, Viral* / blood
  • Communicable Diseases, Emerging / immunology
  • Communicable Diseases, Emerging / prevention & control
  • Communicable Diseases, Emerging / virology
  • False Positive Reactions
  • Hemagglutination Inhibition Tests / methods
  • Hemagglutination Inhibition Tests / statistics & numerical data
  • Humans
  • Influenza A Virus, H5N1 Subtype / genetics
  • Influenza A Virus, H5N1 Subtype / immunology*
  • Influenza Vaccines / immunology*
  • Influenza Vaccines / standards*
  • Influenza, Human / immunology
  • Influenza, Human / prevention & control
  • Influenza, Human / virology
  • Laboratories
  • Neutralization Tests / methods
  • Neutralization Tests / statistics & numerical data
  • Reference Standards
  • Reproducibility of Results
  • Serologic Tests / methods*
  • Serologic Tests / statistics & numerical data
  • Sheep
  • World Health Organization

Substances

  • Antibodies, Viral
  • Influenza Vaccines