牛奶中金黄色葡萄球菌PCR检测方法的建立与应用

被引：8

作者：

吕艳

王华

王君玮

徐天刚

王淑娟

赵永刚

王志亮

机构：

[1] 中国动物卫生与流行病学中心国家外来动物疫病诊断中心ABL-实验室

来源：

现代生物医学进展 | 2009年 / 9卷 / 05期

关键词：

金黄色葡萄球菌; 鉴定; 牛奶; 聚合酶链反应;

D O I：

10.13241/j.cnki.pmb.2009.05.016

中图分类号：

R155 [饮食卫生与食品检查];

学科分类号：

100403 ;

摘要：

目的:建立一种直接从牛奶中检测金黄色葡萄球菌的PCR快速诊断方法。方法:根据已发表的金黄色葡萄球菌16-23SrRNA特异性序列设计并合成一对引物,通过优化反应条件建立从牛奶中直接检测金黄色葡萄球菌的PCR方法。结果:与细菌的常规分离方法相比,PCR法敏感性高,与其它型葡萄球菌无交叉反应,特异性达100%,检测细菌基因组DNA最低浓度为35ng/μL,能在5h内对送检的牛奶样品直接进行金黄色葡萄球菌的测定,可用于乳制品中金黄色葡萄球菌的检测。结论:成功建立了快速检测牛奶中金黄色葡萄球菌PCR方法。

引用

页码：931 / 933

页数：3

共 8 条

[1]

Direct identification of bacteria in clinical respiratory samples using fluorescent amplicon length analysis of 16S–23S rRNA spacer-region[J] . C. Massonet,T. De Baere,J. Van Eldere. &nbspJournal of Microbiological Methods . 2006 (2)

[2]

Technical Note: Improved Method for Rapid DNA Extraction of Mastitis Pathogens Directly from Milk[J] . P. Cremonesi,B. Castiglioni,G. Malferrari,I. Biunno,C. Vimercati,P. Moroni,S. Morandi,M. Luzzana. &nbspJournal of Dairy Science . 2006 (1)

[3] Multiplex polymerase chain reaction assay for simultaneous detection of Staphylococcus aureus and streptococcal causes of bovine mastitis [J].

Phuektes, P ;

Mansell, PD ;

Browning, GF .

JOURNAL OF DAIRY SCIENCE, 2001, 84 (05) :1140-1148

[4] Application of a convenient DNA extraction method and multiplex PCR for the direct detection of Staphylococcus aureus and Yersinia enterocolitica in milk samples [J].

Ramesh, A ;

Padmapriya, BP ;

Chandrashekar, A ;

Varadaraj, MC .

MOLECULAR AND CELLULAR PROBES, 2002, 16 (04) :307-314

[5]

Simultaneous Detection of Mastitis Pathogens, Staphylococcus aureus, Streptococcus uberis, and Streptococcus agalactiae by Multiplex Real-Time Polymerase Chain Reaction[J] . B.E. Gillespie,S.P. Oliver. &nbspJournal of Dairy Science . 2005 (10)

[6] Development of a highly sensitive and specific assay to detect Staphylococcus aureus in bovine mastitic milk [J].

Graber, H. U. ;

Casey, M. G. ;

Naskova, J. ;

Steiner, A. ;

Schaeren, W. .

JOURNAL OF DAIRY SCIENCE, 2007, 90 (10) :4661-4669

[7]

Molecular characterization of Serratia marcescens strains by RFLP and sequencing of PCR‐amplified 16S rDNA and 16S–23S rDNA intergenic spacer[J] . H.Zhu,W.Y.Zhou,M.Xu,Y.L.Shen,D.Z.Wei. &nbspLetters in Applied Microbiology . 2007 (2)

[8] Development of specific and rapid detection of bacterial pathogens in dairy products by PCR [J].

Chotar, M. ;

Vidova, B. ;

Godany, A. .

FOLIA MICROBIOLOGICA, 2006, 51 (06) :639-646

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