CAJ | 학술논문

为建立一种空肠弯曲菌和沙门菌的双重实时荧光定量 PCR 检测方法，根据空肠弯曲菌的保守基因 hipO 和沙门菌的保守基因 invA 分别设计合成引物和 TaqMan 探针，分别使用 FAM、JOE 作为探针报告基团，TAMRA 作为探针淬灭基团。优化反应体系及条件，建立适用于检测食品中空肠弯曲菌和沙门菌的双重实时荧光定量 PCR 方法。结果显示，该检测方法特异性强，灵敏度高，空肠弯曲菌检测限可达10 CFU/mL，沙门菌检测限达230 CFU/mL。表明建立的双重实时荧光定量 PCR 可为实现食品中空肠弯曲菌和沙门菌的同时检测提供新方法。
위건립일충공장만곡균화사문균적쌍중실시형광정량 PCR 검측방법，근거공장만곡균적보수기인 hipO 화사문균적보수기인 invA 분별설계합성인물화 TaqMan 탐침，분별사용 FAM、JOE 작위탐침보고기단，TAMRA 작위탐침쉬멸기단。우화반응체계급조건，건립괄용우검측식품중공장만곡균화사문균적쌍중실시형광정량 PCR 방법。결과현시，해검측방법특이성강，령민도고，공장만곡균검측한가체10 CFU/mL，사문균검측한체230 CFU/mL。표명건립적쌍중실시형광정량 PCR 가위실현식품중공장만곡균화사문균적동시검측제공신방법。
This study aimed to establish a rapid detection of Campylobacter jejuni and Salmonella by duplex real-time PCR.The primers and Taq man probes were designed based on the invA gene of Salmonella and hipO gene of Campylobacter jejuni ,using FAM,JOE and TAMRA fluorescently labeled specific probes aimed at the conserved genes of target pathogens,respectively.We compared Tm values to fix these primers and probes,and optimize the reaction conditions to establish the duplex real-time PCR approach for the detection of Campylobacter jejuni and Salmonella.The sensitivity of the duplex real-time PCR was 10 CFU/mL,230 CFU/mL for Campylobacterjejuni and Salmonella.The duplex real-time PCR showed good specificity which was validated by testing various bacteria isolates.In this study,a duplex PCR with high specificity and sensitivity was developed and it would provide useful information for the simultaneous detection of Campylobacter jejuni and Salmonella in food.