婴幼儿奶粉中阪崎肠杆菌双重荧光PCR快速检测方法的建立  

Novel duplex real-time Taqman PCR assay for rapid detection of Enterobacter sakazakii in powdered infant formula

作　　者：黄建飞[1] 刘小青[1] 刘斌[1] 陈泽峰 兰全学[1] 陈晶[1] HUANG Jian-fei, LIU Xiao-qing, LIU Bin, CHEN Ze-feng, LAN Quan-xue, CHEN Jing(Food Testing Institute, Shenzhen Academy of Metrology ＆ Quality Inspection, Shenzhen 518131, Chin)

机构地区：[1]深圳市计量质量检测研究院,食品检测所,广东深圳518131

出　　处：《食品工业科技》2018年第6期246-249,297,共5页Science and Technology of Food Industry

摘　　要：旨在建立针对阪崎肠杆菌的双重荧光PCR快速检测方法。以阪崎肠杆菌局部大分子合成(MMS)操纵子和外膜蛋白A(ompA)为靶基因,建立双重荧光 PCR 反应体系,探讨该体系的特异性、灵敏度和抗干扰能力。结果表明,双重荧光 PCR 体系对阪崎肠杆菌的灵敏度为4.3×10~3CFU/mL,人工污染初始菌量为2 CFU/100 g奶粉样品增菌24 h即可检出;39株实验菌中的15株阪崎肠杆菌出现特异性扩增,24株非阪崎肠杆菌未出现特异性扩增。本研究所建立的双重荧光PCR体系特异好、灵敏度较高及抗干扰能力强,可用于婴幼儿奶粉中阪崎肠杆菌的快速检测。To develop a dulplex real- time Taqman polymerase chain reaction （ PCR ） assay for the detection of Enterobacter sakazakii.Primers and probes were designed based on the sequences of the macromolecular synthesis （MMS）operon and outer member protein A（ ompA）.Sensitivity and specificity of the assay were evaluated.Detection limit of the assay in pure culture was 4.3 × 103 CFU/mL and as few as 2 CFU/100 g of Enterobacter sakazakii could be detected in artificially contaminated infant formula through 24 h of enrichment. All of 15 Enterobacter sakazakii strains were successfully identified, no specific amplification were presented when 24 non- Enterobacter sakazakii were tested. The duplex real time Taqman PCR assay developed in study was sensitivity, specific and rapid.It could be used for detection of Enterobacter sakazakii from infant milk powder for food safety.

关 键 词：阪崎肠肝菌 荧光PCR ompA基因 MMS基因 

分 类 号：TS201.3[轻工技术与工程—食品科学]