食品中阪崎肠杆菌多重PCR检测方法的建立  

Detection of Enterobacter sakazakii in food by multi- plex PCR

作　　者：楼秀芹[1] 斯国静[1] 戚建江[1] 刘涛[1] 俞骅[1] 

机构地区：[1]杭州市疾病预防控制中心微生物检验科,浙江杭州310021

出　　处：《中国卫生检验杂志》2014年第2期159-163,共5页Chinese Journal of Health Laboratory Technology

基　　金：浙江省杭州市科技局科技发展计划(20110533B19)

摘　　要：目的建立并优化食品中阪崎肠杆菌检测的多重PCR方法。方法以阪崎肠杆菌ITS序列,16s rDNA和ompA基因为靶基因,选择3对引物,建立并优化多重PCR反应体系;验证其特异性、灵敏度和抗干扰能力;并在实际样品中进行阪崎肠杆菌的检测,检测结果与现行国家标准方法进行比较。结果该多重PCR反应体系具有良好的特异性、灵敏度和较强的抗干扰能力。人工污染奶粉样品的模拟实验表明,增菌后该多重PCR反应体系可以检出1 CFU/100 g奶粉的阪崎肠杆菌。108份不同种类的实际样品中共检出阪崎肠杆菌10份,检出率为9.26%,检测结果与采用现行国家标准方法测定具有良好的一致性。结论本研究所建立的多重PCR方法特异好、灵敏较高、结果准确,可用于食品中阪崎肠杆菌的日常检测。Objective To develop a multiplex PCR assay for Enterobacter sakazakii detection in food samples. Methods ITS sequence, 16S rDNA and ompA genes as target genes, three sets of primers were selected to develop a multiplex PCR assay, then its specificity, sensitivity and immunity from interference were verified. This multiplex - PCR assay was used to detect En- terobacter sakazakii in practical samples and the detection results were conpared to those of national standard method. Results The multiplex PCR assay had good performances on specificity, sensitivity and anti - interference. The detection limit of the as- say was 1 CFU/100 g in artificially inoculated infant milk powder samples after the procedure of enrichment. Ten of 108 differ- ent food samples were Enterobacter sakazakii positive, and the positive rate was 9.26%, which was completely consistent with the detection results by GB 4789.40 -2010. Conclusion The method has good specificity, sensitivity and accuracy. It can be used for the routine detection of Enterobacter sakazakii in food.

关 键 词：阪崎肠杆菌 多重PCR 食源性致病菌 

分 类 号：R378.2[医药卫生—病原生物学]