Development of a recombinase-aided amplification combined with a lateral flow dipstick assay for rapid detection of H7 subtype avian influenza virus

作者：Fuyou Zhang, Jiajing Shang, Juan Luo, Xin Yin, Xiaohui Yu, Wenming Jiang, Jinping Li, Liping Yuan, Guangyu Hou, Hualei Liu, Yang Li · 发表于：Frontiers in Microbiology · 年份：2023 · DOI：10.3389/fmicb.2023.1286713 · 被引用次数：8 · 研究领域：Influenza Virus Research Studies、Animal Disease Management and Epidemiology、Respiratory viral infections research

Avian influenza viruses (AIV) pose a significant persistent threat to the public health and safety. It is estimated that there have been over 100 outbreaks caused by various H7 subtypes of avian influenza viruses (AIV-H7) worldwide, resulting in over 33 million deaths of poultry. In this study, we developed a recombinase-aided amplification combined with a lateral flow dipstick assay for the detection of hemagglutinin (HA) genes to provide technical support for rapid clinical detection of AIV-H7. The results showed that the assay can complete the reaction within 30 min at a temperature of 39°C. Specificity tests demonstrated that there was no cross-reactivity with other common poultry pathogens, including Newcastle disease virus (NDV) and infections bronchitis virus (IBV). The detection limit of this assay was 1 × 10 1 copies/μL, while RT-qPCR method was 1 × 10 1 copies/μL, and RT-PCR was 1 × 10 2 copies/μL. The κ value of the RT-RAA-LFD and RT-PCR assay in 132 avian clinical samples was 0.9169 ( p < 0.001). These results indicated that the developed RT-RAA-LFD assay had good specificity, sensitivity, stability and repeatability and may be used for rapid detection of AIV-H7 in clinical diagnosis.