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Multicenter Study
. 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

Karen L Laurie  1 , Othmar G Engelhardt  2 , John Wood  3 , Alan Heath  3 , Jacqueline M Katz  4 , Malik Peiris  5 , Katja Hoschler  6 , Olav Hungnes  7 , Wenqing Zhang  8 , Maria D Van Kerkhove  9 ; CONSISE Laboratory Working Group participants
Collaborators, Affiliations
Multicenter Study

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

Karen L Laurie et al. Clin Vaccine Immunol. 2015 Aug.

Abstract

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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Figures

FIG 1
FIG 1
Reproducibility within laboratories of serology assay results for assays detecting antibodies to A(H1N1)pdm09 (A and B) and A(H3N2) and A(H5N1) (C and D) viruses. Graphs show the (percent) proportions of replicate assays differing by >2-fold (A and C) and >4-fold (B and D) for the 2-day ELISA MN assay and the 3-day MN assay with detection by HA and CPE for each participating laboratory for all sera. ND indicates instances in which the assay or detection method was not performed.
FIG 2
FIG 2
Relationship between test sample titers for antibodies to A(H1N1)pdm09 (A and B), A(H3N2) (C and D), or A(H5N1) (E and F) viruses determined by the 2-day ELISA MN assay and by the 3-day MN assay with detection by HA (A, C, and E) or CPE (B, D, and F). Each laboratory is represented by a color as indicated in the key.
See this image and copyright information in PMC

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