China Animal Health and Epidemiology Center, 369 Nanjing Road, Qingdao, Shandong Province, China.
Yanbian University, Agricultural College, Yanji, Jilin, China.
Virol J. 2022 Jul 30;19(1):129. doi: 10.1186/s12985-022-01807-0.
The H5 subtype avian influenza virus (AIV) has caused huge economic losses to the poultry industry and is a threat to human health. A rapid and simple test is needed to confirm infection in suspected cases during disease outbreaks.
In this study, we developed a reverse transcription recombinase-aided amplification (RT-RAA) assay for the detection of H5 subtype AIV. Assays were performed at a single temperature (39 °C), and the results were obtained within 20 min.
The assay showed no cross-detection with Newcastle disease virus or infectious bronchitis virus. The analytical sensitivity was 10 RNA copies/μL at a 95% confidence interval according to probit regression analysis, with 100% specificity. Compared with published reverse transcription quantitative real-time polymerase chain reaction assays, the κ value of the RT-RAA assay in 420 avian clinical samples was 0.983 (p < 0.001). The sensitivity for avian clinical sample detection was 97.26% (95% CI, 89.56-99.52%), and the specificity was 100% (95% CI, 98.64-100%).
These results indicated that our RT-RAA assay may be a valuable tool for detecting H5 subtype AIV.
H5 亚型禽流感病毒（AIV）给家禽业造成了巨大的经济损失，并且对人类健康构成威胁。在疾病爆发期间，需要一种快速简便的方法来确认疑似病例的感染。
本研究开发了一种用于检测 H5 亚型 AIV 的逆转录重组酶辅助扩增（RT-RAA）检测方法。该检测在单一温度（39°C）下进行，结果可在 20 分钟内获得。
该检测方法与新城疫病毒或传染性支气管炎病毒无交叉检测。根据概率回归分析，分析灵敏度为 95%置信区间内的 10 RNA 拷贝/μL，特异性为 100%。与已发表的逆转录定量实时聚合酶链反应检测方法相比，RT-RAA 检测方法在 420 份禽临床样本中的κ 值为 0.983（p<0.001）。该方法对禽临床样本的检测灵敏度为 97.26%（95%CI，89.56-99.52%），特异性为 100%（95%CI，98.64-100%）。
这些结果表明，我们的 RT-RAA 检测方法可能是检测 H5 亚型 AIV 的一种有价值的工具。
