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Clin Vaccine Immunol. 2015 Aug;22(8):957-64. doi: 10.1128/CVI.00278-15. Epub 2015 Jun 24.

International Laboratory Comparison of Influenza Microneutralization Assays for A(H1N1)pdm09, A(H3N2), and A(H5N1) Influenza Viruses by CONSISE.

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WHO Collaborating Centre for Reference and Research on Influenza, Victorian Infectious Diseases Reference Laboratory, Peter Doherty Institute for Infection and Immunity, Melbourne, Australia
National Institute for Biological Standards and Control, Medicines and Healthcare Products Regulatory Agency, Potters Bar, Hertfordshire, United Kingdom.
National Institute for Biological Standards and Control, Hertfordshire, United Kingdom.
Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
School of Public Health, The University of Hong Kong, Pok Fu Lam, Hong Kong.
Public Health England, London, United Kingdom.
Norwegian Institute of Public Health, Oslo, Norway.
World Health Organization, Geneva, Switzerland.
Center for Global Health, Institut Pasteur, Paris, France.


The microneutralization assay is commonly used to detect antibodies to influenza virus, and multiple protocols are used worldwide. These protocols differ in the incubation time of the assay as well as in the order of specific steps, and even within protocols there are often further adjustments in individual laboratories. The impact these protocol variations have on influenza serology data is unclear. Thus, a laboratory comparison of the 2-day enzyme-linked immunosorbent assay (ELISA) and 3-day hemagglutination (HA) microneutralization (MN) protocols, using A(H1N1)pdm09, A(H3N2), and A(H5N1) viruses, was performed by the CONSISE Laboratory Working Group. Individual laboratories performed both assay protocols, on multiple occasions, using different serum panels. Thirteen laboratories from around the world participated. Within each laboratory, serum sample titers for the different assay protocols were compared between assays to determine the sensitivity of each assay and were compared between replicates to assess the reproducibility of each protocol for each laboratory. There was good correlation of the results obtained using the two assay protocols in most laboratories, indicating that these assays may be interchangeable for detecting antibodies to the influenza A viruses included in this study. Importantly, participating laboratories have aligned their methodologies to the CONSISE consensus 2-day ELISA and 3-day HA MN assay protocols to enable better correlation of these assays in the future.

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