滚环扩增法检测副溶血性弧菌  

Detection of Vibrio parahaemolyticus With Rolling Circle Amplification Technique

作　　者：张健[1,2] 姜英辉[2] 管斌[1] 雷质文[2] 房保海[2] 唐静[2] 梁成珠[2] 

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛266000 [2]山东出入境检验检疫局,山东青岛266000

出　　处：《海洋科学进展》2011年第A01期144-149,共6页Advances in Marine Science

基　　金：山东出入境检验检疫局研究项目--滚环扩增和荧光偏振技术同步检测食源性致病菌及其标准化研究（SK208821）

摘　　要：为建立快速准确的副溶血性弧菌的检测体系，根据滚环扩增方法的原理设计引物、锁式探针以及反应体系。锁式探针两端与目标序列特异性结合，并用DNA连接酶使其形成环状，然后利用引物滚环扩增环形核酸，最后用凝胶电泳方法验证扩增产物。试验结果显示，该方法可以成功检测副溶血性弧菌，并不会扩增其他菌，灵敏度要比普通PCR方法高。研究结果显示，RCA检测体系具有很好的特异性、灵敏度和稳定性，适用于副溶血性弧菌的检测和鉴定。To establish a new rapid and accurate method to detect Vibrio parahaernolyticus, principle of rolling circle amplification （RCA）technique was used to design primers, padlock probe and reaction system. The padlock probe consists of two target complementary regions at 5 ＇ and 3＇ end, it will form a ring under the function of DNA ligase, then use probe rolling circle to amplify nucleotide and verify products with gel electrophoresis. The result showed that the method can detect Vibrio parahaernolyticus specifically, and the detection sensitivity of RCA was higher than that of conventional PCR. It proved that RCA is a capable approach to detect Vibrio parahaemolyticus. The method is specific, stable and reliable.

关 键 词：滚环扩增 副溶血性弧菌 锁式探针 

分 类 号：Q78[生物学—分子生物学]