CAJ | 학술논문

通过结合两种技术-免疫磁分离技术与双重荧光定量PCR技术,建立同时、快速对鲜猪肉进行沙门氏菌和金黄色葡萄球菌的检测方法.利用偶联有特异性抗体免疫磁球,在37℃的条件下能够有效地从250 mL循环体系中边富集边捕获目标菌.利用特异性的引物与探针,对两种食源性致病菌进行双重荧光定量PCR同时检测.本研究方法针对沙门氏菌和金黄色葡萄球菌的检测限分别达到3.6 CFU/g和9.2 CFU/g.方法总体灵敏度、特异性和准确度达到98.8%、100%以及98.9%.本研究建立的免疫磁分离-双重荧光定量PCR方法比国标方法更快速和高效,适用于鲜猪肉中沙门氏菌和金黄色葡萄球菌的快速检测.
통과결합량충기술-면역자분리기술여쌍중형광정량PCR기술,건립동시、쾌속대선저육진행사문씨균화금황색포도구균적검측방법.이용우련유특이성항체면역자구,재37℃적조건하능구유효지종250 mL순배체계중변부집변포획목표균.이용특이성적인물여탐침,대량충식원성치병균진행쌍중형광정량PCR동시검측.본연구방법침대사문씨균화금황색포도구균적검측한분별체도3.6 CFU/g화9.2 CFU/g.방법총체령민도、특이성화준학도체도98.8%、100%이급98.9%.본연구건립적면역자분리-쌍중형광정량PCR방법비국표방법경쾌속화고효,괄용우선저육중사문씨균화금황색포도구균적쾌속검측.
In this study,a rapid and simultaneous detection for Salmonella sp.and Staphylococcus (S.) aureus from fresh pork samples was developed by combining immunomagnetic separation (IMS) with duplex real-time PCR (RT-PCR).Magnetic beads coated with specific antibodies were used to capture the pathogens from 250 mL at 37℃.Then,duplex RT-PCR was applied with two sets of specific primers and probes.The limit of detections were 3.6 CFU/g for Salmonella,and 9.2 CFU/g for S.aureus.The sensitivity,specificity,and accuracy of IMS-multiplex RT-PCR method were 98.8%,100%,and 98.8%,respectively.IMS-duplex RT-PCR method is more rapid and effective than GB method,and which was suitable for the rapid detection of Salmonella and Staphylococcus aureus in fresh pork.