xTAG液相芯片技术检测多种食源性致病菌方法的研究  

Development of a Luminex xTAG Assay for the Rapid Detection of Common Food Borne Pathogens

作　　者：彭延[1] 余波[1] 张静玲[1] 邹文菁[1] PENG Yan;YU Bo;ZHANG Jing-ling;ZOU Wen-jing(Hubei Provincial Center for Disease Control and Prevention,Wuhan,Hubei 430079,China)

机构地区：[1]湖北省疾病预防控制中心,湖北武汉430079

出　　处：《公共卫生与预防医学》2022年第5期54-56,共3页Journal of Public Health and Preventive Medicine

基　　金：湖北省卫生计生委2018年度第四批联合基金立项项目(WJ2018H254)。

摘　　要：目的建立一种基于xTAG微球标记技术,同时快速检测4种食源性致病菌的方法。方法分别针对沙门氏菌的invA基因、副溶血性弧菌的toxR基因、单核细胞增生李斯特菌的hly基因、金黄色葡萄球菌的Sa442基因,设计标记有MagTAG微球Taq的引物,建立多重PCR实验方法,将PCR产物与带有反向Taq序列的微球杂交,使用Luminex200检测仪检测杂交结果,并对检测方法的灵敏度、特异性进行评价。结果该方法能同时检出4种食源性致病菌,特异性好。副溶血性弧菌toxR、单核细胞增生李斯特菌hly、沙门氏菌invA检测的灵敏度为100CFU/mL;金黄色葡萄球菌Sa442的灵敏度为1×10^(3)CFU/mL。结论此研究建立的方法能够快速、准确、特异性的同时检测沙门氏菌、副溶血性弧菌、单核细胞增生李斯特菌、金黄色葡萄球菌。Objective To develop a bead-based xTAG assay for the rapid detection of Salmonella,Vibrio parahaemolyticus,Listeria monocytogenes and Staphylococcus aureus.Methods Specific primers with xTAG were designed and synthesized based on the target gene sequences and MagPlex-TAG Microspheres.After a multiplex PCR amplification,the PCR products hybridized with MagPlex-TAG Microspheres.The Luminex200 array system was used to detect the results of hybridization.The sensitivity and specificity of the method was valued.Results The Luminex xTAG Assay was developed for a specific,rapid,accurate detection of Salmonella,Vibrio parahaemolyticus,Listeria monocytogenes and Staphylococcus aureus.The sensitivity of detection of toxR gene of Salmonella,hly gene of Vibrio parahaemolyticus,and invA gene of Listeria monocytogenes was 100 CFU/mL.The sensitivity of detection of Sa442 gene of Staphylococcus aureus was 1×10^(3)CFU/mL.Conclusion The Luminex xTAG Assay could rapidly,accurately and specifically detect four foodborne pathogens Salmonella,Vibrio parahaemolyticus,Listeria monocytogenesand,Staphylococcus aureus.

关 键 词：液相芯片 食源性致病菌 MagTAG微球 LUMINEX 

分 类 号：R155[医药卫生—营养与食品卫生学]