摘要
目的对Allglo探针和Taqman探针检测GI型和GII型诺如病毒的双重荧光RT-PCR方法进行比较,并应用于临床标本的检测。方法设计针对GI型和GII型诺如病毒的Allglo探针和Taqman探针,优化最佳反应条件,建立一步法荧光RT-PCR快速检测反应体系。对这两种方法的特异性、灵敏度进行比较评估,对56份临床粪便标本进行检测,并统计结果。结果这两种方法特异性强;最低检测限均为102copies/μL以下;但是Allglo探针在同一浓度下比Taqman探针CT值更低,扩增效率更高。对56份临床粪便标本进行检测,两种方法结果一致。结论 Allglo探针双重荧光RT-PCR方法较之Taqman探针检测GI型和GII型诺如病毒扩增效率更高,两者均可用于感染性腹泻暴发中诺如病毒的快速筛查。
Objective To compare Allglo probe and Taqman probe in detection of norovirus genogroup Ⅰ and Ⅱ by double fluorescent RT -PCR method. Methods Taqman probe and Allglo probe were designed according to norovirus genogronp GⅠ and GⅡ . The reaction conditions were optimized to establish the one - step fluorescent RT - PCR rapid detection system . Then the specificity and sensitivity of the two methods were compared. In addition, 56 clinical specimens were detected by the two methods and the results were analyzed by statistics. Results The results showed both the two methods had high specificity. The detection limits of the two methods for norovirus GⅠ and G Ⅱ detection were both below 102 copies/μL. But at the same concentration, Allglo probe has lower CT value and higher amplification efficiency than the Taqman probe. The two methods showed consistent results in detection of 56 clinical specimens. Conclusion Compared to Taqman probe, Allglo probe has higher amplification efficiency in double fluorescent RT - PCR detection of norovirus GⅠ and GⅡ , both can be used for rapid screening of infectious diarrhea outbreaks of norovirus.
出处
《中国卫生检验杂志》
北大核心
2013年第14期2924-2926,2947,共4页
Chinese Journal of Health Laboratory Technology
基金
江苏出入境检验检疫局科技计划项目(2010KJ17)
