乳中志贺氏菌的荧光定量PCR检测方法的建立  

Establishment of Real-time PCR Method for Detection of Shigella in Milk

作　　者：郭瑜[1] 姚笛[1] 侯婷婷[1] 张微[1] 佐兆杭 刘鑫[1] 郭德军[1] 

机构地区：[1]黑龙江八一农垦大学食品学院,黑龙江大庆163319

出　　处：《食品研究与开发》2016年第10期127-130,共4页Food Research and Development

基　　金：黑龙江省大学生创业项目(HNK11A-10-03201410223014)

摘　　要：为了快速、有效的检测乳中的志贺氏菌,建立一种特异性强、灵敏度高的荧光定量PCR方法。根据GenBank公布的志贺氏菌ipaH基因的保守序列设计特异性引物,对提取的志贺氏菌DNA模板进行PCR扩增,对目的基因进行克隆和测序,然后利用荧光定量PCR方法,对志贺氏菌进行检测,确定其扩增条件。建立的方法特异性强,检测的灵敏度可达到10-6 ng/μL。利用建立的方法可检测乳中2.84 cfu/mL的志贺氏菌。该方法为快速、准确检测乳中的志贺氏菌提供了参考。For fast, effective detection of Shigella in milk, a strong specificity, high sensitivity real-time PCR method was established. The specific primers were designed according to the conserved sequence of Shigella i-paH gene published in GenBank, the extracted Shigella DNA template was amplified by PCR, the target gene was cloned and sequenced, then using the fluorescent quantitative PCR method, to detect Shigella, determine the amplification conditions. The established method specificity is high, the detected sensitivity reach 10-6 ng/μL. Using the method of Shigella in milk can be detected at 28.4 cfu/mL. The method provides a reference for fast, accurate detection Shigella in milk.

关 键 词：荧光定量PCR 志贺氏菌 IPAH基因 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]