反转录重组酶辅助扩增检测 H5 亚型禽流感病毒。
Reverse transcription recombinase-aided amplification assay for H5 subtype avian influenza virus.
机构信息
China Animal Health and Epidemiology Center, 369 Nanjing Road, Qingdao, Shandong Province, China.
Yanbian University, Agricultural College, Yanji, Jilin, China.
出版信息
Virol J. 2022 Jul 30;19(1):129. doi: 10.1186/s12985-022-01807-0.
BACKGROUND
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.
METHODS
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.
RESULTS
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%).
CONCLUSIONS
These results indicated that our RT-RAA assay may be a valuable tool for detecting H5 subtype AIV.
背景
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。
方法
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。
结果
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%）。
结论
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。
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