布鲁氏菌多重聚合酶链反应鉴定研究

刘国栋; 崔步云; 刘荣臻; 李元凯

doi:10.3784/j.issn.1003-9961.2008.5.271

布鲁氏菌多重聚合酶链反应鉴定研究

doi: 10.3784/j.issn.1003-9961.2008.5.271

1.
山西医科大学，太原 030009;
2.
中国疾病预防控制中心传染病预防控制所;
3.
山西医科大学汾阳学院医学检验系

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出版历程
- 收稿日期: 2008-04-29
- 刊出日期: 2008-05-30

Shanxi Medical University, Taiyuan 030009, China

1.
Shanxi Medical University, Taiyuan 030009, China

摘要

摘要: 目的　在同一体系中对布鲁氏菌6个种进行鉴定。方法　根据布鲁氏菌6个种的差异基因设计5对引物，应用多重聚合酶链反应（PCR）方法对布鲁氏菌进行鉴定。先用标准菌株建立方法，优化实验条件，然后扩增地方株进行验证。结果　除绵羊附睾种外多重引物PCR方法能将其余5个布鲁氏菌种全部鉴定出来。结论　这种方法快速准确，且对实验操作人员安全，是布鲁氏菌鉴定的辅助方法。
- 布鲁氏菌 /
- 多重聚合酶链反应 /
- 鉴定
Abstract: Objective　The study was conducted to distinguish the six species of Brucella in a single system using multiplex polymerase chain reaction(PCR). Methods　Five pairs of primers were designed according to the gene differences among the six species of Brucella. Mixture?鄄system based PCR was then conducted to identify the strains. Standard Brucella strains were used to establish the process and optimize experimental conditions. Wild strains were cloned for cross verification. Results　Multiplex PCR was capable of identifying each species of Brucella except for B. ovis. Conclusion Multiplex PCR can serve as a quick, accurate and safe supplementary method for the identification of Brucella.
- Brucella /
- multiplex polymerase chain reaction /
- identification

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参考文献(1)

[1]

［1］ Matyas Z, Fujikura T. Brucellosis as a world problem ［J］. Dev Biol Stand, 1984, 56：3-20. ［2］ Garcia?鄄Yoldi D, Marin CM, Miguel MJ, et al. Multiplex PCR assay for the identification and differentiation of all Brucella species and the vaccine strains Brucella abortus S19 and RB51 and Brucella melitensis Rev1［J］. Clin Chem, 2006, 52（4）：779-781. ［3］ Halling SM, Peterson?鄄Burch BD, Bricker BJ, et al. Completion of the genome sequence of Brucella abortus and comparison to the highly similar genomes of Brucella melitensis and Brucella suis. J Bacteriol［J］. 2005，187：2715-2726. ［4］ Rajashekara G, Glasner JD, Glover DA, et al. Comparative whole-genome hybridization reveals genomic islands in Brucella species ［Ｊ］. J Bacteriol, 2004, 186：5040-5051. ［5］ Bricker BJ, Halling SM. Differentiation of Brucella abortus bv.1,2,and 4, Brucella melitensis, Brucella ovis, and Brucella suis bv 1 by PCR［Ｊ］. Clin Microbiol, 1994, 32：2660-2666. ［６］ Ocampo?鄄Sosa AA, Aguero-Balbin J, Garcia?鄄Lobo JM. Development of a new PCR assay to identify Brucella abortus biovars 5, 6 and 9 and the new subgroup 3b of biovar 3［J］. Veter Microbiol， 2005，110: 41-51. ［７］ Cui BY. Brucella pulse field gel electrophoresi typing research ［D］. Shanxi Medical universty, master paper, 2006. （in Chinese）崔步云. 布鲁氏菌脉冲场凝胶电泳分型研究［D］. 山西医科大学，硕士论文，2006. ［８］ Cloeckaert A, Verger JM, Grayon M, et al. Restriction site polymorphism of the genes encoding the major 25 kDa and 36 kDa outer-membrane proteins of Brucella［M］. Microbiology, 1995, 141: 2111-2121. ［９］ Le Fleche P， Jacques I， Grayon M，et al. Evaluation and selection of tandem repeat loci for a Brucella MLVA typing assy［Ｊ］. BMC Microbiol, 2006, 6：9 （1）: 9.

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文章访问数: 1149
HTML全文浏览量: 15
PDF下载量: 393
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布鲁氏菌多重聚合酶链反应鉴定研究

doi: 10.3784/j.issn.1003-9961.2008.5.271

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Shanxi Medical University, Taiyuan 030009, China

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