奶粉中阪崎肠杆菌PCR检测方法研究.pdfVIP

INSPECTION AND QUARANTINE SCIENCE Vol. 15 No.4 2005 4 研究报告 PCR 高旗利 张 霞 罗茂凰 张海滨 张海英 姚 霞 张宏伟 刘 寅 黄熙泰 (, ,300456) ( , , 300071) : 建立和提出奶粉中阪崎肠杆菌PCR 检测方法 利用细菌16S 和23S rDNA 的保守区 作为通用 物, 对6 株阪崎肠杆菌16S~ 23S rDNA 间区序列( ISR) 进行了扩增和测序, 在比较阪崎肠杆菌与其近源 株16S1~ 23S rDNA 间区序列的基础上, 设计了11 条阪崎肠杆菌PCR 检测 物, 组合成30 对PCR 物并筛选出一 对阪崎肠杆菌PCR 检测的特异性 物, 建立了奶粉中阪崎肠杆菌PCR 检测方法 用10 株阪崎肠杆菌, 18 株近源菌验证试验表明, 本文所建立的PCR 方法特异性强; 加菌试验表明, 奶粉样品中阪崎肠杆菌检测低限为 2.2~ 5.4cfu/ 100g, 灵敏度高; 新建的PCR 方法与FDA AM 方法比较试验表明, 两种方法的检测结果完全符合 本文提出的奶粉中阪崎肠杆菌PCR 检测方法填补了国内空白, 达到了国际先进水平, 可在实际工作中推广 : 奶粉; 阪崎肠杆菌;PCR DETECTION OF Enterobacter sakazak ii FROM DEHYDRATED POWDERED MILK BY PCR Gao Qili,Zhang Xia,Luo Maohuang,Zhang Haibin Zhang Haiying,Yao Xia,Zhang Hongwei. Liu Yin,Huang Xitai (Tianjin Exit- Entry Inspection and Quarantine ureau,Tianjin, 300456) (Tianjin Nankai University,Tianjin,300071) Abstract: [Objective] To establish a PCR methodfor detection of E nterobacter sakaz akii in dehydrated powderedmilk. [Method] The conserved sequences in 16S rRNA and 23S rRNA genes of bacteriawere used as a pair of universal primers to amplify the 16S- 23S interspacer region of 6 strains of Enter obacter sakaz akii . On the basis of comparative sequence analysis of the 16S- 23S interspacer regions, 11 primers were designed and30 pairsof these primerswere combined so that a pair of primers specific for E nterobacter sakaz akii was screened out. Even- tually, we established the PCR method for detection of dehydrated powdered milk from dehydrated powdered milk. [ Res lt] The PCR method established in this article is highly specific and sensitive for detection of Enter obacter sakazakii through the testing of 10 strains of En- terobacter sakazakii and 18 strains of other relative bacteria and ar