| 用四重PCR检测方法检测产单核细胞李斯特菌 |
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| 引用本文： | 葛菲菲,徐锋,邱亚峰,沈莉萍,王建.用四重PCR检测方法检测产单核细胞李斯特菌[J].动物医学进展,2010,31(12). |
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| 作者姓名： | 葛菲菲 徐锋 邱亚峰 沈莉萍 王建 |
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| 作者单位： | 1. 上海市动物疫病预防控制中心,上海,201103
2. 中国农业科学院上海兽医研究所,上海,200241 |
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| 基金项目： | 国家科技支撑计划项目(2009BADB9B01) |
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| 摘 要： | 为了同时快速准确的检测产单核细胞李斯特菌以及是否为有毒力的产单核细胞李斯特菌,根据相关文献报道的四重PCR方法,针对该菌的种特异性基因inl A基因进行引物设计,扩增片段大小为800bp,结果应用该PCR法可特异性的扩增出目的基因片段,与GenBank上发表的序列同源性为97.3%。同时通过对毒力基因inlB,inlC和inlJ的检测用来判断菌株的相关毒力。而同一属的其他菌种无害李斯特菌(L.innocua),韦氏李斯特菌(L.welshimeri)均无特异条带,嗜水气单胞菌(J-1),产气荚膜梭菌(C57-13),大肠埃希菌(ATCC 25922),鼠伤寒沙门菌(C79-32),金黄色葡萄球菌(ATCC 25923)结果均为阴性。对临床上送检的23份样品进行四重PCR方法检测并同时与国家标准GB/T22429-2008食品中单核细胞增生李斯特氏菌的快速筛选检验中使用的检测方法进行比对,两种检测方法的结果完全符合,均检测出7份阳性样品。并且四重PCR法表明这7份阳性样品中的产单核细胞李斯特菌均携带有3种毒力因子(inlB,inlC和inlJ),因此该四重PCR法可用于快速鉴定是否是有毒力的产单核细胞李斯特菌,为有效预防控制产单核细胞李斯特菌污染提供技术支撑。
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| 关 键 词： | 产单核细胞李斯特菌 四重PCR 检测 |
Establishment of Species-and Virulence-specific Detection of L.monocytogenes by Quadruple Polymerase Chain Reaction |
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| GE Fei-fei,XU Feng,QIU Ya-feng,SHENG Li-ping,WANG Jian.Establishment of Species-and Virulence-specific Detection of L.monocytogenes by Quadruple Polymerase Chain Reaction[J].Progress In Veterinary Medicine,2010,31(12). |
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| Authors: | GE Fei-fei XU Feng QIU Ya-feng SHENG Li-ping WANG Jian |
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| Institution: | GE Fei-fei1,XU Feng1,QIU Ya-feng2,SHENG Li-ping1,WANG Jian1(1.Shanghai Animal Disease Control Center,Shanghai,201103,China,2.Shanghai Veterinary Research Institute,Chinese Academy of Agricultural Sciences,200241,China) |
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| Abstract: | In order to detect Listeria monocytogenes and its virulence,PCR primers of species-specific inlA gene were designed and the amplified product was 800 bp according to the quadruple PCR.The homology of amplified gene fragment and the published sequence in GenBank was 97.3%.At the same time,the virulence of these strains was ascertained by the formation of 884 bp and / or 517 bp and/or 238 bp fragments with the inlB,inlC and inlJ primers,respectively.However,no PCR products were detected from L.innocua,L.welsh... |
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| Keywords: | Listeria monocytogenes quadruple PCR detection |
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