【摘要】 目的建立和评价一种适合国境口岸及基层检验部门使用的沙门氏菌快速检测方法,在2010年上海世博会保障期间开展应用研究。方法针对沙门氏菌agfA基因,运用环介导等温扩增技术(loop mediated isothermalamplification,LAMP),设计引物进行检测,在65℃恒温条件下、60 min特异性扩增,荧光显色观察结果。对建立的方法进行特异性、灵敏度试验及模拟样本灵敏度试验,与荧光定量PCR、胶体金方法进行比对。并对54株临床分离阳性样本加以验证。结果沙门氏菌属6种不同血清型菌株LAMP检测都呈阳性,其他4种肠道细菌均为阴性;LAMP法的灵敏度与RT-PCR方法接近,达103CFU/ml,是胶体金检测法的100～1 000倍;54株沙门氏菌临床分离样本实验符合率达到100%。结论 LAMP技术能够在简易的实验条件下快速报告结果,检测成本较低、特异性高、灵敏度高,适合口岸从业人员及基层检验部门进行沙门氏菌的初步筛查工作。更多还原

【Abstract】 Objective To establish and evaluate a rapid method for detecting Salmonella at ports and basic clinical laboratory.Methods A set of primers were designed to amplify the salmonella agfA gene under 65℃ for 60 min by LAMP method.The specificity and sensibility of LAMP were evaluated by comparing with RT-PCR and colloid gold immunity chromatography assay.Results All LAMP reactions of 6 serotypes of salmonella were positive,and others were negative.The detection limit of LAMP was 103 CFU /ml which was close to RT-PCR and was 100~1000 times of colloid gold immunity chromatography assay.The results of 54 bacterial strains amplified by LAMP were all positive and the compliance rate was 100%.Conclusion The LAMP assay established in this study is a sensitive,rapid and simple tool for detecting Salmonella without expensive apparatus.Thus,it could be used as a primary screen method to detect Salmonella of employees at ports and basic clinical laboratory.更多还原