Evaluation of a proximity extension assay for the detection of H1 2009 pandemic influenza viruses

Abstract The rapid influenza diagnostic tests (RIDTs) are widely distributed, simple to use, but often lack sensitivity as compared to gold standard methods (viral culture and nucleic acid detection technologies). Applying RIDTs outside of epidemic or pandemic infections results in large numbers of false negatives. Hence, a sensitive RIDT that would reduce the number of false negatives would result in an increased clinical value. We evaluated the potential of a proximity extension assay (PEA) for the detection of influenza A H1 viruses. This technology makes use of antibodies to capture the pathogen, followed by molecular detection. Forty-seven nasopharyngeal swab samples, all confirmed infections of the H1 2009 pandemic influenza virus, were evaluated. The performance of PEA was compared to the RIDT Quickvue Influenza A + B assay. The success rate of the comparative assays was modeled by means of a binary logistic response model. Both assays performed equally well within the current range of viral particles, expressed as log 10 copies/ml. When the actual input of viral particles was taken into account, the 95% hitrate of PEA lies within the range of 4.60–7.02 log 10 copies/reaction, which is an almost 2 log 10 sensitivity improvement over the 95% hitrate of the Quickvue RIDT, ranging from 6.86 to 9.37 log 10 copies/reaction. The PEA method holds promise to improve sensitive detection of influenza viruses in clinical samples.