Changchun Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Changchun, China.
College of Animal Medicine, Jilin University, Changchun, China.
Front Cell Infect Microbiol. 2023 Jan 5;12:1071288. doi: 10.3389/fcimb.2022.1071288. eCollection 2022.
Influenza A viruses (IAVs) are important pathogens of respiratory infections, causing not only seasonal influenza but also influenza pandemics and posing a global threat to public health. IAVs infection spreads rapidly, widely, and across species, causing huge losses, especially zoonotic IAVs infections that are more harmful. Fast and sensitive detection of IAVs is critical for controlling the spread of this disease.
Here, a real-time reverse transcription recombinase-aided amplification (real-time RT-RAA) assay targeting conserved positions in the matrix protein gene (M gene) of IAVs, is successfully established to detect IAVs. The assay can be completed within 20 min at 42°C.
The sensitivity of the real-time RT-RAA assay was 142 copies per reaction at 95% probability, which was comparable to the sensitivity of the RT-qPCR assay. The specificity assay showed that the real-time RT-RAA assay was specific to IAVs, and there was no cross-reactivity with other important viruses. In addition, 100%concordance between the real-time RT-RAA and RT-qPCR assays was achieved after testing 120 clinical specimens.
The results suggested that the real-time RT-RAA assay we developed was a specific, sensitive and reliable diagnostic tool for the rapid detection of IAVs.
甲型流感病毒（IAV）是呼吸道感染的重要病原体，不仅会引发季节性流感，还会引发流感大流行，对全球公共卫生构成威胁。IAV 感染传播迅速、广泛且跨物种，造成巨大损失，特别是具有人畜共患性的 IAV 感染更为严重。快速、灵敏地检测 IAV 对于控制该疾病的传播至关重要。
本研究建立了一种基于甲型流感病毒基质蛋白基因（M 基因）保守区的实时逆转录重组酶辅助扩增（real-time RT-RAA）检测方法，用于检测 IAV。该方法在 42°C 下可在 20 分钟内完成。
实时 RT-RAA 检测的灵敏度为 95%概率时 142 拷贝/反应，与 RT-qPCR 检测的灵敏度相当。特异性检测表明，实时 RT-RAA 检测方法特异性针对 IAV，与其他重要病毒无交叉反应。此外，对 120 份临床标本进行检测，实时 RT-RAA 与 RT-qPCR 检测方法的一致性达到 100%。
研究结果表明，我们开发的实时 RT-RAA 检测方法是一种快速检测 IAV 的特异性、灵敏性和可靠的诊断工具。
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