CAJ | 학술논문

目的探讨多色组合探针编码技术实时PCR(MCPC-PCR)检测食品标本中的沙门菌和金黄色葡萄球菌等病原细菌的检测限,并应用于食物中毒标本的检测.方法配制系列浓度(101～109CFU/ml)的菌悬液,以鲜肉样品和蛋糕制备模拟样本各11份,应用MCPC-PCR方法检测22份模拟标本中沙门菌和金黄色葡萄球菌;101份冻存的食物中毒标本营养肉汤增菌液,以及某次食物中毒事件的5例首批患者肛拭营养肉汤增菌液提取DNA后进行MCPC-PCR检测.结果 MCPC-PCR技术在食品标本中沙门菌和金黄色葡萄球菌检测的检测限为102copies/test(每个PCR测试管含有的拷贝数);101份冻存的食物中毒标本营养肉汤增菌液以MCPC-PCR检测,结果阳性23份,其中18份经细菌学方法分离菌株证实,MCPC-PCR和细菌学方法检测结果均相同的为96份,结果符合率达95.05%;采自某次食物中毒事件的5个首批患者肛拭标本MCPC-PCR检测结果提示为副溶血性弧菌,与细菌学分离结果完全符合.结论MCPC-PCR技术灵敏度高、特异度好,可应用于多种食源性病原菌的快速筛查和快速检测.
목적 탐토다색조합탐침편마기술실시PCR(MCPC-PCR)검측식품표본중적사문균화금황색포도구균등병원세균적검측한,병응용우식물중독표본적검측.방법 배제계렬농도(101～109CFU/ml)적균현액,이선육양품화단고제비모의양본각11빈,응용MCPC-PCR방법검측22빈모의표본중사문균화금황색포도구균;101빈동존적식물중독표본영양육탕증균액,이급모차식물중독사건적5례수비환자항식영양육탕증균액제취DNA후진행MCPC-PCR검측.결과 MCPC-PCR기술재식품표본중사문균화금황색포도구균검측적검측한위102copies/test(매개PCR측시관함유적고패수);101빈동존적식물중독표본영양육탕증균액이MCPC-PCR검측,결과양성23빈,기중18빈경세균학방법분리균주증실,MCPC-PCR화세균학방법검측결과균상동적위96빈,결과부합솔체95.05%;채자모차식물중독사건적5개수비환자항식표본MCPC-PCR검측결과제시위부용혈성호균,여세균학분리결과완전부합.결론MCPC-PCR기술령민도고、특이도호,가응용우다충식원성병원균적쾌속사사화쾌속검측.
Objective To investigate the detection limit of multicolor combinational probe coding real-time PCR (MCPC-PCR) in detection of Salmonella and Staphylococcus aureus suspended in the food samples, and to apply MCPC-PCR to detect the samples of food poisoning. Methods Series concentration of bacterium suspension (101 - 109 CFU/ml) was prepared by using 22 simulated samples including fresh meat and cakes and then MCPC-PCR was applied to detect Salmonella and Staphylococcus aureus in 22 samples. Enrichment broth of 101 frozen samples and 5 early patients' anal swabs in food poisoning cases were detected after the DNA samples were extracted. Results The limits of MCPC-PCR assay in detecting Salmonella and Staphylococcus aureus were about 102 copies/test; 101 frozen enrichment broth of samples in food poisoning cases were detected by MCPC-PCR assay, of 23 positive samples, 18 were confirmed by bacteriology techniques; 96 samples detected by MCPC-PCR and bacteriology techniques had the same results,and the coincidence rate was 95.05%. Anal swabs, collected from 5 of early patients in a food poisoning case gave a clue to be Vibrio parahaemolyticus by MCPC-PCR assay and then were perfectly consistent with bacteriology assay. Conclusion As a method of high sensitivity and good specificity, MCPC-PCR assay can quickly and conveniently detect multiple pathogens existing in food samples, therefore we recommend it to be used in rapidly screening or simultaneous detection of food-berne diseases.