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Influenza Other Respir Viruses. 2015 Jun 13. doi: 10.1111/irv.12333. [Epub ahead of print]
Optimization of a micro-neutralisation assay and its application in antigenic characterisation of influenza viruses.
Lin Y1, Gu Y1, Wharton SA1, Whittaker L1, Gregory V1, Li X1, Metin S1, Cattle N1, Daniels RS1, Hay AJ1, McCauley JW1.
Author information
Abstract
BACKGROUND:
The identification of antigenic variants and the selection of influenza viruses for vaccine production are based largely on antigenic characterisation of the haemagglutinin (HA) of circulating viruses using the haemagglutination inhibition (HI) assay. However, additional to evolution related to escape from host immunity, variants emerging as a result of propagation in different cell substrates can complicate interpretation of HI results.
OBJECTIVES:
The objective was to develop further a micro-neutralisation (MN) assay to complement the HI assay in antigenic characterisation of influenza viruses to assess the emergence of new antigenic variants and reinforce the selection of vaccine viruses.
METHODS:
A 96-well-plate plaque reduction MN assay based on measurement of the Infected Cell Population (ICP) using a simple imaging technique.
RESULTS:
Improvements to the plaque reduction MN assay included selection of the most suitable cell line according to virus type or subtype, and optimisation of experimental design and data quantitation. Comparisons of the results of MN and HI assays showed the importance of complementary data in determining the true antigenic relationships among recent human influenza A(H1N1)pdm09, A(H3N2) and type B viruses.
CONCLUSIONS:
Our study demonstrates that the improved MN assay has certain advantages over the HI assay: it is not significantly influenced by the cell-selected amino acid substitutions in the neuraminidase (NA) of A(H3N2) viruses, and it is particularly useful for antigenic characterisation of viruses which either grow to low HA titre and/or undergo an abortive infection resulting in an inability to form plaques in cultured cells. This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.
KEYWORDS:
Influenza; antigenicity; haemagglutination inhibition; micro-neutralisation
PMID: 26073976 [PubMed - as supplied by publisher]
Optimization of a micro-neutralisation assay and its application in antigenic characterisation of influenza viruses.
Lin Y1, Gu Y1, Wharton SA1, Whittaker L1, Gregory V1, Li X1, Metin S1, Cattle N1, Daniels RS1, Hay AJ1, McCauley JW1.
Author information
Abstract
BACKGROUND:
The identification of antigenic variants and the selection of influenza viruses for vaccine production are based largely on antigenic characterisation of the haemagglutinin (HA) of circulating viruses using the haemagglutination inhibition (HI) assay. However, additional to evolution related to escape from host immunity, variants emerging as a result of propagation in different cell substrates can complicate interpretation of HI results.
OBJECTIVES:
The objective was to develop further a micro-neutralisation (MN) assay to complement the HI assay in antigenic characterisation of influenza viruses to assess the emergence of new antigenic variants and reinforce the selection of vaccine viruses.
METHODS:
A 96-well-plate plaque reduction MN assay based on measurement of the Infected Cell Population (ICP) using a simple imaging technique.
RESULTS:
Improvements to the plaque reduction MN assay included selection of the most suitable cell line according to virus type or subtype, and optimisation of experimental design and data quantitation. Comparisons of the results of MN and HI assays showed the importance of complementary data in determining the true antigenic relationships among recent human influenza A(H1N1)pdm09, A(H3N2) and type B viruses.
CONCLUSIONS:
Our study demonstrates that the improved MN assay has certain advantages over the HI assay: it is not significantly influenced by the cell-selected amino acid substitutions in the neuraminidase (NA) of A(H3N2) viruses, and it is particularly useful for antigenic characterisation of viruses which either grow to low HA titre and/or undergo an abortive infection resulting in an inability to form plaques in cultured cells. This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.
KEYWORDS:
Influenza; antigenicity; haemagglutination inhibition; micro-neutralisation
PMID: 26073976 [PubMed - as supplied by publisher]