CAJ | 학술논문

根据沙门氏菌invA基因、副溶血性弧菌toxR基因和大肠杆菌O157：H7 RFBE基因的保守序列，设计引物和探针，通过优化反应体系，测定其灵敏度和特异性，建立了可同时检测上述三种致病菌的多重实时荧光PCR方法。该方法对纯茵的检测灵敏度均低于1O cfu／PCR反应体系。人工染菌样品经6h增菌，检测的灵敏度可低于10cfu／25g；对48株标准／参考菌株的检测结果，只有目的菌株呈阳性反应；而定量检测批内和批间的变异系数均小于2％。应用本法检测人工染菌样品和进出口水产品样品，结果与sN标准方法检测结果一致。本法可在8h内完成对水产品中上述三种致病菌的定性或定量检测，可适应于进出口水产品的快速检验。
근거사문씨균invA기인、부용혈성호균toxR기인화대장간균O157：H7 RFBE기인적보수서렬，설계인물화탐침，통과우화반응체계，측정기령민도화특이성，건립료가동시검측상술삼충치병균적다중실시형광PCR방법。해방법대순인적검측령민도균저우1O cfu／PCR반응체계。인공염균양품경6h증균，검측적령민도가저우10cfu／25g；대48주표준／삼고균주적검측결과，지유목적균주정양성반응；이정량검측비내화비간적변이계수균소우2％。응용본법검측인공염균양품화진출구수산품양품，결과여sN표준방법검측결과일치。본법가재8h내완성대수산품중상술삼충치병균적정성혹정량검측，가괄응우진출구수산품적쾌속검험。
A multiplex real-time PCR method was developed for the simultaneous detection of Salmonella spp, Vibrio parahaemolytichus and Escherichia Coli O157：H7 based on the primers and probes designed based on the conserved domains of invA gene for Salmonella spp, toxR gene for Vibrio parahaemolytichus and RFBE gene for Escherichia Coli O157： }t7- The developed multiplex real-time PCR method could detect less than 10efu per PCR reaction for all the pathogenic bacteria above in pure cultured broth and less than 10cfu per 25g for 6h-cultured artificially inoculated samples. Only target strains of 48 reference/standard strains were detected positive in the specificity testing. The intra- and inter-assay coefficients of variation were less than 2%. The method was applied to samples artificially coniaminated with these pathogenic bacteria and samples collected from aquatic products for entry and exit. The detection results were consistent with those of SN reference methods. It took less than 8h to detect these three pathogenic bacteria qualitatively or quantitatively in samples of aquatic products, and it could be applied to the quick detection of these three pathogenic bacteria in aquatic products for entry and exit.