摘要
文章旨在建立一种可同时快速检测食品中沙门氏菌和副溶血弧菌的TaqMan双重荧光定量聚合酶链式反应(Polymerase chain reaction,PCR)检测方法。分别针对沙门氏菌的invA基因和副溶血弧菌的tlh基因序列的保守片段,设计了特异性检测引物和探针,并优化其使用浓度及反应退火温度,建立双重荧光定量PCR检测体系,并对方法的灵敏度、特异性及稳定性进行评估。结果显示:建立的双重荧光定量PCR检测法可特异性扩增沙门氏菌和副溶血弧菌,其他菌株无扩增。沙门氏菌检测灵敏度最低检测值可达1 copies/μL,副溶血弧菌检测灵敏度为10 copies/μL。批内批间变异系数均<2%,重复性和稳定性较好,与传统方法检测结果一致,检测时长大幅度缩短。综上表明,建立的双重荧光PCR检测方法能够快速、准确地检测出食品中沙门氏菌和副溶血弧菌,为食品中食源性致病菌的快速检测提供技术支持。
This study aims to establish a TaqMan dual fluorescent quantitative polymerase chain reaction(PCR)detection method for the rapid simultaneous detection of Salmonella and Vibrio parahaemolyticus in food.Specific detection primers and probes targeting the conservative sequences of the invA gene of Salmonella and the tlh gene of Vibrio parahaemolyticus were designed,and their concentrations and annealing temperatures were optimized to establish a dual-fluorescent quantitative PCR detection system,and evaluating the sensitivity,specificity and stability of the method,The results showed that the established dual fluorescence quantitative PCR detection method could specifically amplify Salmonella and Vibrio parahaemolyticus,while other strains did not amplify.The minimum detection value of Salmonella detection can reach 1 copies/μL,and the sensitivity of Vibrio parahaemolyticus detection is 10 copies/μL.The intra-batch and inter-batch coefficients of variation were both less than 2%,and the repeatability and stability were good,the results were consistent with those of traditional methods,and the time is shortened.In summary,the dual fluorescent PCR detection method established can quickly and accurately detect Salmonella and Vibrio parahaemolyticus in food,which can provide technical support for the rapid detection of foodborne pathogens bacteria in food.
作者
康俊杰
王秋悦
吴晨雨
苑中策
董洁琼
杨鹏
陈秀英
郑百芹
KANG Junjie;WANG Qiuyue;WU Chenyu;YUAN Zhongce;DONG Jieqiong;YANG Peng;CHEN Xiuying;ZHENG Baiqin(Tangshan Food and Drug Comprehensive Testing Center,Tangshan 063000,China;Key Laboratory of Preventive Veterinary Medicine in Hebei Province,College of Animal Science&Technology,Hebei Normal University of Science&Technology,Qinhuangdao 066000,China;Hunchun Animal Husbandry Station,Hunchun 133300,China)
出处
《食品科技》
CAS
北大核心
2024年第3期351-356,共6页
Food Science and Technology
基金
河北省第四批高端人才项目
唐山市科技创新领军人才项目(21130243A)
河北省青年拔尖人才项目
河北省预防兽医学重点实验室绩效后补助项目(20567621H)。
作者简介
康俊杰(1985-),女,高级兽医师,研究方向为农产品兽药残留检验;通信作者:陈秀英;通信作者:郑百芹。
