RT-LAMP is a potential future molecular diagnostic tool for influenza A virus

Plain language summaryInfluenza A virus (IAV) is responsible for severe disease worldwide and causes significant mortality and morbidity. A sensitive, specific tool for the detection of IAV infection in its early stages is needed, so we designed an HNB-based RT-LAMP assay for IAV detection. This is an easy, cost-effective, sensitive, and specific test that may become a point-of-care test for IAV detection during outbreaks. The limit of detection (LOD) was found to be 10 fg/ml in a purified strain of H1N1/H3N2 by our LAMP assay, which was ten-times more sensitive than conventional RT-PCR/M-RT-PCR diagnostic tests. Aim: Influenza A virus (IAV) causes serious illness and is responsible for significant morbidity and mortality. To diagnose IAV infection in its early stages, a quick, sensitive, precise detection method is needed for effective clinical management. Materials & methods: In-house hydroxylnaphthol blue (HNB)-based RT-LAMP assay for early detection of IAV using the HA gene was compared with RT-PCR/multiplex-RT-PCR. Results: For the reference strains of IAV, (H1N1 (A/Texas/50/2012) and H3N2 (A/Malaysia/2089302/2009)) RT-LAMP and RT-PCR/M-RT-PCR exhibited a limit of detection (LOD) of 10 and 100 fg/ml, respectively. Conclusion: HNB-based RT-LAMP is a rapid, sensitive, cost-effective diagnostic tool, and could be a point-of-care test for IAV patients during outbreaks. Tweetable abstractAn LOD of 10 fg/ml was exhibited by both agarose and HNB-based RT-LAMP, while a LOD of 100 fg/ml was obtained by conventional RT-PCR/M-RT-PCR assay.