Impact of influenza virus escape‐mutations on influenza detection by the rapid influenza diagnostic test

Abstract

During the influenza pandemic of 2009-2010, rapid influenza diagnostic tests (RIDTs) were used to detect influenza viral infections because they are quick and simple to use. However, retrospective studies showed that RIDTs performed poorly when used to diagnose pandemic viral infections. Determining how amino acid sequence changes in pandemic or epidemic influenza viral antigens impact clinical value of RIDTs has not been possible, because the viral epitopes recognized by RIDTs have been not mapped. In this study, the effect of escape-variations or mutations in influenza viral antigens upon the sensitivity and specificity of an RIDT was investigated by characterizing the immunological properties of the antibodies used in the RIDT. Escape-mutants were generated by cultivating A/Korea/01/2009 in the presence of an excess of the same antibodies used in the RIDT. Escape-mutants not recognized by the RIDT were selected. Epitopes recognized by the RIDT were mapped by comparing the sequence and immunological analysis of the escape-variants and wild-type isolates. The RIDT antibodies recognized epitopes on the Sa antigenic site and in the F subdomain in hemagglutinin. Variants bearing mutations in these epitopes were not detected by the RIDT. The frequency of escape-variants emerging since the 2009-2010 pandemic was calculated as 1.27% using in silico surveillance of influenza sequence databases. These results suggest that mapping the relevant epitopes of RIDTs and making such information available to clinics would be helpful for determining whether RIDTs match newly emergent strains and subtypes prior to retrospective re-evaluation of the RIDTs using clinical specimens.