Rapid detection of human influenza A viruses by HFman probe-based loop-mediated isothermal amplification assays

Since China abandoned the zero-COVID policy at the end of 2022, a wave of severe Flu pandemic emerged in China. Rapid and accurate diagnosis of Influenza A virus (IAV) is critical for clinical management and therapeutic decision-making of patients with fever. Here, we reported a novel IAV HF-LAMP assay, which can be performed with purified RNA or directly using clinical samples. The assays with purified RNA and clinical samples have high sensitivity with limit of detection (LOD) of 9.6 copies/reaction, 9900 copies/mL, and short sample-to-answer times of 36 and 50 min, respectively. Both assays showed high specificity and significantly higher IAV detection rate than the rapid antigen detection (RAD) assays. Furthermore, we found the vast majority (91.2 %) of children with fever during the pandemic were infected by IAV, and current IAV infection has a very narrow detectable window. The novel IVA HF-LAMP assays will provide robust tools to facilitate early diagnosis of IAV infection in current and future seasonal influenza epidemics. •An IAV HF-LAMP assay with high sensitivity was developed.•The extraction-free format of the assay has a short sample-to-answer time.•The vast majority (91.2 %) of children with fever were infected by IAV.•Current IAV infection has a very narrow detectable window.•A duplex assay for detection of both IAV and SARS-CoV-2 in a single tube was developed.