黄热病毒逆转录环介导等温扩增检测方法的建立

Establishment of a reserve transcription loop-mediated isothermal amplification detection method for Yellow fever virus

【目的】建立一种简便快速的黄热病毒分子检测方法。【方法】运用RT-LAMP(Reserve transcription loop-mediated isothermal amplification)技术设计一套引物特异性识别黄热病毒E基因中的6个区域,建立高效简便的黄热病毒等温扩增检测方法。【结果】该检测方法具有良好的特异性,与其他虫媒病毒无交叉反应。同时检测灵敏度可达到0.066 pg,与RT-PCR相比,灵敏度高100倍左右。模拟样本检测与预期相符,临床样本未检测到阳性标本。【结论】该方法为黄热病毒检疫提供了一种可在简便环境中特异、灵敏的快速检测手段。

[Objective] Establish a simple and rapid molecular detection of Yellow fever virus. [Methods] The reserve transcription loop-mediated isothermal amplification （RT-LAMP） that amplifies RNA with high specificity and rapidity under an isothermal condition was applied for rapid detection of Yellow fever virus. A set of primers were designed specifically to identify six special areas of Yellow fever virus’s E gene. [Results] The results indicated that the method had high specificity, and no cross amplification occurred with other arbovirus RNAs. The sensitivity of the RT-LAMP assay was 0.066 pg, which was 100 times higher than that of RT-PCR. The detection results of simulation samples were consistent with expected. [Conclusion] The established rapid detection method for Yellow fever virus that can be carried out in a simple experimental environment has been proved to be specific and sensitive.