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Position: Home > Articles > Comparative Study of Detection of Norovirus in Oysters by RT-PCR,RT-Seminested PCR and RT-LAMP FOOD SCIENCE 2009,30 (12) 246-250

三种分子生物学方法检测牡蛎中诺如病毒的比较研究

作  者：
李丹;唐庆娟;李兆杰;王玉明;薛勇;薛长湖
单  位：
中国海洋大学食品科学与工程学院
关键词：
诺如病毒;牡蛎;检测;RT-PCR;RT-半巢式PCR;RT-LAMP
摘  要：
采用RT-PCR、RT-半巢式PCR(seminested PCR)和RT-环介导等温扩增(LAMP)三种分子生物学方法分别检测了牡蛎中经粪便污染的诺如病毒。三种方法所采用的特异性引物均针对诺如病毒高度保守的N/S结构域。结果显示:一步法RT-PCR较两步法结果理想但仍不能有效去除食品中的PCR反应抑制物。RT-半巢式PCR和RT-LAMP的特异性和敏感性都远远优于RT-PCR,但是RT-半巢式PCR操作繁琐费时。RT-LAMP扩增程序简单、反应时间短,且不需要精密的温度循环装置,在产物中加入SYBR Green Ⅰ染料后可用肉眼直接判断反应结果。因此,RT-LAMP有望发展成为快速检测牡蛎中诺如病毒的有效手段。
译  名：
Comparative Study of Detection of Norovirus in Oysters by RT-PCR,RT-Seminested PCR and RT-LAMP
作  者：
LI Dan,TANG Qing-juan,LI Zhao-jie,WANG Yu-ming,XUE Yong,XUE Chang-hu(College of Food Science and Engineering,Ocean University of China,Qingdao 266003,China)
关键词：
norovirus;oyster;detection;RT-PCR;RT-seminested PCR;RT-LAMP
摘  要：
Reverse transcription polymerase chain reaction(RT-PCR),RT-seminested PCR and reverse transcription loop-mediated isothermal amplification(RT-LAMP) assays were compared to detect norovirus in oysters.All the three assays targeted the highly conserved capsid N-terminal/shell(N/S) domain of norovirus with specific primers.Although one-step RT-PCR presented more apparent results than two-step approach,it still had the limitation in eliminating inhibitors contained in food matrix,and both RT-seminested PCR and RT-LAMP showed significantly higher specificity and sensitivity than RT-PCR.However,the operation of RT-seminested PCR is tedious and time-consuming.In comparison to other two approaches,the amplification program of RT-LAMP is simple,and the reaction time is short.Without special apparatus needed,RT-LAMP can be performed under isothermal condition and the results may be visually ascertained by a simple color reaction using SYBR Green Ⅰdye.Therefore,RT-LAMP is of great potential to become a valuable method for rapid detection of norovirus in oysters.

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