Data_Sheet_1_Development of a recombinase-aided amplification combined with a lateral flow dipstick assay for rapid detection of H7 subtype avian influenza virus.ZIP

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 × 101 copies/μL, while RT-qPCR method was 1 × 101 copies/μL, and RT-PCR was 1 × 102 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.

禽流感病毒（Avian influenza viruses，AIV）对公共卫生安全构成持续且严峻的威胁。据统计，全球范围内已发生逾100起由不同H7亚型禽流感病毒（AIV-H7）引发的疫情，造成超3300万只家禽死亡。本研究开发了一种重组酶介导扩增（recombinase-aided amplification，RAA）联合侧流层析试纸条检测（lateral flow dipstick assay，LFD）的方法，用于血凝素（hemagglutinin，HA）基因的检测，以期为AIV-H7的临床快速检测提供技术支撑。实验结果显示，该检测方法可在39℃条件下30分钟内完成反应。特异性验证实验表明，该方法与其他常见家禽病原体无交叉反应，包括新城疫病毒（Newcastle disease virus，NDV）与传染性支气管炎病毒（Infectious Bronchitis Virus，IBV）。本方法的检测限为1×10¹拷贝/μL；实时荧光定量反转录聚合酶链式反应（RT-qPCR）的检测限为1×10¹拷贝/μL，而反转录聚合酶链式反应（RT-PCR）的检测限为1×10²拷贝/μL。在132份家禽临床样本中，反转录重组酶介导扩增-侧流层析试纸条检测（RT-RAA-LFD）与RT-PCR方法的κ值为0.9169（p<0.001）。上述结果表明，本研究开发的RT-RAA-LFD检测方法具备良好的特异性、灵敏度、稳定性与重复性，可用于AIV-H7的临床快速诊断与检测。