Development and application of real-time PCR assays for specific detection of contemporary avian influenza virus subtypes N5, N6, N7, N8 and N9
2018
Previously-published neuraminidase (NA) subtype-specific reverse transcriptase RealTime (RRT)-PCRs were further validated for the detection of five avian influenza virus (AIV) NA subtypes, namely N5, N6, N7, N8 and N9. Testing of 30 AIV isolates of all nine NA subtypes informed the assay assessments, with the N5 and N9 RRT-PCRs retained as the original published assays, while the N7 and N8 assays were modified in the primer / probe sequences to optimise detection of current threats. The preferred N6 RRT-PCR was either the original or modified variant, depending on the specific H5N6 lineage. Clinical specimen (n=137) testing revealed the selected N5, N6 and N8 RRT-PCRs to sensitively detect clade 2.3.4.4b highly-pathogenic (HP)AIV infections due to H5N5, H5N6 and H5N8 subtypes respectively, all originating from European poultry and wild bird cases during 2016-2018. Similar testing (n=32 clinical specimens) also showed the N7 and N9 RRT-PCRs to sensitively detect European H7N7 HPAIV and China-origin H7N9 lo...
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