Absolute Quantification of H5-Subtype Avian Influenza Viruses Using Droplet Digital Loop-Mediated Isothermal Amplification

Human infection= with avian influenza A HSN1-viruses can cause severe diseases with high mortality rate and, continues to pose a significant threat to global public health. Rapid diagnosis is needed for identifying the types of influenza viruses for making timely treatment decisions. Here, we demonstrate absolute quantification of HS-subtype influenza viruses-by digital loop-thediated isothermal amplification (dLAMP) on our recently-developed cross-interface emulsification (XiE) method. Our results show that XiE-based dLAMP is, highly specific and displays comparable sensitivity to real-time PCR (qPCR) and digital FCR. (dPCR). Notably,.dLAMP is more tolerant to inhibitOry substanCes than PCR methods and demonstrated similar detection efficiency to qPCR for real _HSN1,samples. Therefore, it can serve as a robust and precise alternative to qPCR or dPCR and is especially suitable for en-vironmentaLand-clinical-samples with hard-to-remove contaminants. We believe that our dLAMP method offers great potential for rapid and :accurate diagnosis of influenza, and other infectious diseases.