动物源性食品中志贺氏菌实时荧光定量PCR快速检测方法的建立  

Development of a dual real- time PCR for the rapid detection of Shigella in animal-origined food

作　　者：李丹丹[1] 徐义刚 王绥家[1] 高慎阳[3] 李一经[4] 

机构地区：[1]海南出入境检验检疫局检验检疫技术中心,海口570311 [2]黑龙江出入境检验检疫局检验检疫技术中心,哈尔滨150001 [3]辽宁医学院畜牧兽医学院,辽宁锦州121001 [4]东北农业大学动物医学学院,哈尔滨150030

出　　处：《东北农业大学学报》2014年第8期98-102,共5页Journal of Northeast Agricultural University

基　　金：海南省自然科学基金项目(310106);国家质检总局科技项目(2013IK031);国家质检总局科技项目(2012IK157);海南省应用技术研究与开发专项项目(ZDXM20130025)

摘　　要：根据志贺氏菌属高度保守的ipaH基因序列,设计探针和引物,通过优化反应条件,建立检测动物源性食品中志贺氏菌实时荧光定量PCR方法,应用于动物源性食品中志贺氏菌的快速检验。结果表明,该法灵敏度约为2.8 cfu·mL-1,经对205份肉类、蛋、奶及其制品和动物腹泻物、人工污染样品等进行检测,共检出13份阳性样本,与国标(GB 4789.5-2012)方法的检测结果一致。表明建立的荧光PCR方法操作简便、特异性强、灵敏度高,具有良好实用性。According Shigella ipaH highly conserved gene sequences, probes and primers designed by optimizing the reaction conditions, the establishment of foods of animal origin Shigella real-time PCR method, used in foods of animal origin Shiga rapid test for Shigella. The results showed that the sensitivity of a dual real-time PCR was 2.8 cfu·mL-1. 13 from 205 samples of meat, egg, milk and itsproducts, animal diarrhea materials and artificial contamination samples were positive in a dual real- time PCR assay, which was in accordance with the testing result according to GB 4789.5-2012.The results showed that a dual real- time PCR assay developed in this work was simple, specificity, high sensitivity, good practicality for the detection of Shigella.

关 键 词：志贺氏菌 IPAH基因 荧光定量PCR 快速检测 

分 类 号：S855.12[农业科学—临床兽医学]