银白色葡萄球菌实时荧光PCR快速检测方法的建立  

Development of real-time PCR for the rapid detection of Staphylococcus argenteus

作　　者：郭正洋 刘钟栋[1] 王远洋 陈晶[2] 陈国培[2] 兰全学[2] 刘小青[2] GUO Zheng-yang1, LIU Zhong-dong1, WANG Yuan-yangt, CHEN Jing2 , CHEN Guo- pei2 , LAN Quan- xue2, LIU Xiao- qing2＇ ＊(1. Henan University of Technology, Zhengzhou 450001, China ; 2.Shenzhen Academy of Metrology ＆ Quality Inspection, Shenzhen 518131, Chin)

机构地区：[1]河南工业大学,河南郑州450001 [2]深圳市计量质量检测研究院,广东深圳518131

出　　处：《食品工业科技》2018年第6期222-226,共5页Science and Technology of Food Industry

基　　金：深圳市科技计划项目(JSGG20160606144217004);国家重点研发计划资助项目(2016YFD0400800

摘　　要：本文旨在建立一种银白色葡萄球菌实时荧光 PCR 检测方法。根据对 NCBI 数据库中银白色葡萄球菌非核糖体多肽合成酶(NRPS)基因序列的比对分析,设计引物及探针,并基于所设计的引物及探针建立 Taq-man 实时荧光 PCR 的检测方法。结果表明,本实验所建立的方法特异性较强,只能扩增出银白色葡萄球菌,而其他常见菌株的检测结果呈阴性;该方法的灵敏度为10 pg/uL;在对实验室通过传统方法分离的金黄色葡萄球菌的检测中,发现有两个分离菌株呈阳性,其结果与普通 PCR 方法完全一致。In this study, a real time polymerase chain reaction assay for detection of newly emerged staphylococcus argenteus was established.Based on the result of muhialignment analysis on the published staphylococcus argenteus NRPS sequences on NCBI database, a pair of primers specific and probe were designed, and then a RT-PCR for detection of staphylococcus argenteus was established by utilizing the designed primers and probe.The results showed that detection of common pathogenic bacterias ,only staphylococcus argenteus was positive, indicating the established RT-PCR assay was highly specific.The detection limit of 10 pg/uL of the assay was determined.In the laboratory separation of Staphylococcus aureus detection ,two positive isolates were found,the results was in complete accord with ordinary PCR method.

关 键 词：银白色葡萄球菌 非核糖体多肽合成酶 实时荧光PCR 

分 类 号：TS201.3[轻工技术与工程—食品科学]