Avian influenza A viruses (AIV) significantly impact both animal and human health, necessitating reliable diagnostics for effective control, particularly for highly pathogenic strains such as H5Nx. This study details the development and laboratory validation of the Alveo Sense Poultry Avian Influenza Tests, designed for rapid, on-site detection of the AIV M-gene and subtypes H5, H7, and H9 using reverse-transcription loop-mediated isothermal amplification (RT-LAMP) and impedance-based measurements. These tests, using single-use microfluidic cartridges, deliver results within 45 minutes. 

Laboratory validation demonstrated 100% specificity for AIV subtypes without cross-reactivity with non-AIV pathogens. The tests showed a limit of detection (LoD95) for H5, H7, and H9 subtypes ranging between RT-PCR Ct values of 29–33 in cloacal and oropharyngeal samples and could detect AIV in both spiked and field samples. Reproducibility and repeatability studies confirmed perfect agreement across operators and laboratories, maintaining accuracy under different pre-analytical conditions. These tests provide rapid, accurate, and reliable diagnostics for AIV subtypes, valuable for early detection and outbreak control, with further field validation nee