实时逆转录环介导等温扩增法快速检测GI型诺如病毒

Rapid determination of norovirus genogroup I by real-time reverse transcription loop-mediated isothermal amplification

中文关键词:  GI型诺如病毒  实时  逆转录环介导等温扩增法

英文关键词:norovirus genogroup I  real-time  reverse transcription loop-mediated isothermal amplification

中文摘要:

      目的 建立实时逆转录环介导等温扩增法(reverse transcription loop-mediated isothermal amplification, RT-LAMP)快速检测GI型诺如病毒的分析方法。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、样品检测应用等, 进行GI型诺如病毒实时RT-LAPM法检测关键技术的研究。结果 针对GI型诺如病毒特异性基因, 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GI型诺如病毒。最佳检测温度为65 ℃, 最小检测限为22.6拷贝/μL, 且只对GI型诺如病毒靶基因特异, 检测应用结果可靠。结论 本研究建立了GI型诺如病毒的实时RT-LAMP检测方法, 可用于GI型诺如病毒的样品检测。

英文摘要:

      Objective To establish a method for rapid determination of norovirus genogroup I by real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) method. Methods Through the selection of reaction primers, optimization of detection temperature, study of sensitivity and specificity, and application of sample detection, the key technology of real-time rt-lapm for detection of norovirus genogroup I was studied. Results Aiming at the specific gene of norovirus genogroup I, the best primer group was selected by LAMP method, and the primer group was used to detect norovirus genogroup I specifically, sensitively and quickly. The optimum detection temperature was 65 ℃, the minimum detection limit was 22.6 copies/μL, it was only specific to norovirus genogroup I, and the test results were reliable. Conclusion A real-time RT-LAMP method for rapid detecting norovirus genogroup I is established, which can be used for clinical detection of norovirus genogroup I.