| 沙门菌和志贺菌二重PCR检测方法的建立及应用 |
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| 引用本文： | 盘宝进,罗兆飞,韦梅良,汪文龙,刘军义,陈立标.沙门菌和志贺菌二重PCR检测方法的建立及应用[J].动物医学进展,2009,30(5). |
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| 作者姓名： | 盘宝进 罗兆飞 韦梅良 汪文龙 刘军义 陈立标 |
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| 作者单位： | 1. 广西出入境检验检疫局技术中心,广西南宁,530021
2. 东兴出入境检验检疫局,广西东兴,535651 |
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| 基金项目： | 国家质检总局科技兴检项目 |
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| 摘 要： | 根据GenBank提供的沙门菌invA基因序列和志贺菌ipaH基因序列设计引物,建立了二重PCR方法,在同一反应体系同时检测两种致病菌核酸,经二重PCR方法扩增后,在同一泳道同时检出沙门菌284 bp和志贺菌611 bp特异性扩增条带,而普通大肠埃希菌、金黄色葡萄球菌、变形杆菌、阪畸大肠埃希菌、绿脓杆菌、蜡样芽胞杆菌、马链球菌、产单核细胞李斯特菌、鹅大肠埃希菌、猪水肿病大肠埃希菌均未出现这两种条带.对PCR产物进行测序,测序结果与已发表的基因核苷酸序列比较,沙门菌同源性为93%～100%,志贺菌同源性为98%～100%.应用建立的沙门菌和志贺菌二重PCR方法对广西6个试验猴养殖场1 665份猴粪便样品进行检测,检出沙门菌阳性25份,志贺菌阳性90份,阳性检出率分别为1.5%和5.4%.表明建立的沙门菌和志贺菌二重PCR检测方法特异性强、敏感性高,适用于临床粪便样品的快速检测.
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| 关 键 词： | 猴 沙门菌 志贺菌 二重聚合酶链式反应 检测 |
Establishment and Application of Double PCR Detection Method for Salmonella and Shigella |
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| PAN Bao-jin,LUO Zhao-fei,WEI Mei-liang,WANG Wen-long,LIU Jun-yi,CHEN Li-biao.Establishment and Application of Double PCR Detection Method for Salmonella and Shigella[J].Progress In Veterinary Medicine,2009,30(5). |
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| Authors: | PAN Bao-jin LUO Zhao-fei WEI Mei-liang WANG Wen-long LIU Jun-yi CHEN Li-biao |
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| Institution: | 1.Technology Centre of Guangxi Entry-Exit Inspection and Quarantine Bureau;Nanning;Guangxi;530021;China;2.Dongxing Entry-Exit Inspection and Quarantine Bureau;Dongxing;535651;China |
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| Abstract: | Two pair of specific primers for the polymerase chain reaction(PCR) according to the relevant nucleotide sequences of Salmonella invA and Shigella ipaH in GenBank were designed.Double PCR method was established to detect DNA of Salmonella and Shigella in one reaction system.A 284bp DNA fragment of Salmonella and a 611 bp of Shigella were found,while the nagtive strains were not found.The sequences of amplified fragments were detected and compared with all relevant genes.The nucleotide homologies of Salmonel... |
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| Keywords: | simian Salmonella Shigella double PCR detection |
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