利用滚环扩增技术检测食源性致病微生物的-研究和应用.pdfVIP

个温度下即可进行，最终得到所需的扩增序列，利用核酸电泳技术，观察到核酸 的长度大小，据此判断扩增是否成功，如果扩增成功即证明核酸样品中含有目标 菌的特异性序列，即证明检测的样品是阳性。 研究结果显示本课题构建的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、单核增生李斯特氏菌、志贺氏菌和小肠结肠炎耶尔 森氏菌检测和鉴定。并研制成可以应用的试剂盒。用试剂盒与传统方法相比较后 发现，其具有如下几个优势：检测时间短、灵敏度高、准确性优异、试剂盒携带 方便。 关键词：滚环扩增， 副溶血性弧菌， 单核细胞增生李斯特氏菌，志贺氏菌，小 肠结肠炎耶尔森氏菌 Detection of food-borne pathogens with Rolling Circle Amplification Technique Abstract Food-borne pathogens are pathogens and bacteria can cause illness in human with food as the transmission medium. In this paper, the focus are Vibrio parahaemolyticus, Listeria monocytogenes,Shigella and Yersinia enterocolitica. The traditional method of detection of food-borne pathogens is culture method. The procedure is preliminary enrichment culture, isolation and purification with selective medium, and identification by biochemical test. The method is cumbersome ,with a longer time. The accuracy and sensitivity is not very good. In order to solve this problem, Nucleotide amplification technique was used in this experiment. The specific sequence of the target bacteria was amplified, and was electrophoresed by the gel electrophoresis , to make sure that the size of amplication bands were the same as that of target bacteria sequence . At present, the mature method of nucleotide amplification is the PCR amplification technique, but the PCR amplification technique also has some drawbacks. The PCR nucleotide amplification technique requires that the target sequence must be switched between several temperature, so the PCR instrument is requisite and expensive. Rolling circle amplification techniq