基于toxR基因的PCR检测副溶血性弧菌的方法建立  

Development of a toxR-based PCR assay for detecting Vibrio parahaemolyticus.

作　　者：黄晨阳[1] 于龙[1] 兰智杰[1] 任烁[1] 李振宁[1] 王晓东[1] 

机构地区：[1]海军总医院海军卫生防疫队,北京101113

出　　处：《河南预防医学杂志》2013年第5期327-330,共4页Henan Journal of Preventive Medicine

基　　金：军队课题(AHJ10C001)

摘　　要：目的建立基于toxR基因的PCR检测副溶血弧菌的方法,并通过实验条件摸索,优化反应体系条件,验证PCR方法检测副溶血弧菌的优势。方法应用基于toxR基因的环引物PCR法和已报道的PCR法对副溶血性弧菌和食品样品进行检测,并优化反应条件。对两种检测方法的灵敏度、特异度和检测时间进行比较。结果环引物PCR法可以使副溶血性弧菌扩增出大小180 bp的toxR基因片段,两种方法检测副溶血性弧菌的特异度均为100%,检出限分别为4×103CFU/mL和5×104CFU/mL,检测所需时间分别为2 h 50 min和3 h10 min。结论基于toxR基因的环引物PCR法检测副溶血性弧菌的特异性更强,检出限更低,检测所需时间更短,能够较好地满足卫生防疫机构快速检测副溶血性弧菌的需要。Objective To develop a toxR-based PCR assay for detecting Vibrio parahaemolyticus,by exploring and optimizing conditons of test to verify the advantages of PCR on detecting Vibrio parahaemolyticus at the same time.Methods Vibrio parahaemolyticus and food samples were tested by using toxR-based PCR assay and PCR assay,optimized the conditons of test.The sensitivity,specificity and testing time of toxR-based PCR assay were compared with PCR assay.Results a size of 180 bp gene was amplified by toxR-based PCR assay,the specificity of both assay were 100 %,detection limit was 4×103CFU/mL and 5×104CFU/mL respectively,testing time of both assay was 2 h50 min and 3 h10 min respectively.Conclusion toxR-based PCR assay was better in high specificity,low detection and less reaction time,which made it better satisfied the need of rapid detection of Vibrio parahaemolyticus for health and epidemic prevention agencies.

关 键 词：副溶血性弧菌 PCR toxR基因 

分 类 号：R123.5[医药卫生—环境卫生学]