实时逆转录环介导等温扩增法快速检测甲型肝炎病毒

Rapid determination of hepatitis A virus by real-time reverse transcription loop-mediated isothermal amplification

中文关键词:  甲型肝炎病毒  实时  逆转录环介导等温扩增法  快速检测

英文关键词:hepatitis A virus  real-time  reverse transcription loop-mediated isothermal amplification  rapid detection

中文摘要:

      目的 建立甲型肝炎病毒(hepatitis A virus, HAV)实时逆转录环介导等温扩增(reverse transcription loop-mediated isothermal amplification, RT-LAMP)快速检测方法。方法 通过反应引物的筛选、检测温度的优化、灵敏度和特异性研究、模拟样品检测应用, 进行甲型肝炎病毒实时RT-LAMP法检测体系建立。结果 针对甲型肝炎病毒特异性基因, 通过设计筛选出最佳实时RT-LAMP引物组, 并使用该组引物特异、灵敏、快速地检测到甲型肝炎病毒。最佳检测温度为63 ℃, 最小检测限为10拷贝/μL, 且只对甲型肝炎病毒靶基因特异, 检测应用结果可靠。结论 实时RT-LAMP技术可以用于食品中甲型肝炎病毒的高效特异性检测, 为食品中甲型肝炎病毒的检测工作提供更加有效的技术保障。

英文摘要:

      Objective To establish a rapid detection method for hepatitis A virus (HAV) by real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP). Methods The real-time RT-LAMP detection system for hepatitis A virus was established by screening reaction primers, optimizing the detection temperature, studying the sensitivity and specificity, and detecting the application of simulated samples. Results Aiming at the hepatitis A virus specific gene, the optimal real-time RT-LAMP primer group was screened out by design, and the hepatitis A virus was detected specifically, sensitively and rapidly by using the primer group. The optimal detection temperature was 63 ℃, and the minimum detection limit was 10 copies/μL, moreover, it was specific only for the target gene of hepatitis A virus, so the detection application result was reliable. Conclusion Real-time RT-LAMP technology can be used for efficient and specific detection of hepatitis A virus in food, which provides more effective technical support for the detection of hepatitis A virus in food.