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布鲁氏菌病病原快速检测方法的建立 被引量:5

A rapid detection method for pathogen of brucellosis
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摘要 根据编码牛种布鲁氏菌外膜蛋白(OMP-2)基因的核苷酸序列设计1对PCR引物,并对PCR扩增条件进行优化,建立了快速检测布鲁氏菌病病原的PCR方法。特异性检测表明,该引物对牛种、羊种、猪种布鲁氏菌制备的模板DNA均能扩增出193bp目的基因片段,但不能扩增大肠杆菌、金黄色葡萄球菌、大肠结肠炎耶氏菌(0∶9型)的目的基因片段;敏感性检测显示,乳样的检测灵敏度达50cfu/mL、纯化的布鲁氏菌DNA检测灵敏度达1.5pg,可重复性和稳定性十分理想。可见,该布鲁氏菌病病原PCR检测方法具有较高的特异性和敏感性,能为快速诊断布鲁氏菌病提供技术支撑。 Based on the nucleotide sequence encoding OMP-2 gene of Brucella abrtus, a pair of PCR primers were designed and the amplification conditions of PCR were optimized to establish the rapid detection method for the pathogen of brucellosis. The specific detection results showed that 193bp gene fragment could be amplified in template DNA of Brueella abrtus, Brueella melltensis and Brucella suis, the pathogens of brucellosis, by the specific primers, while it could not he amplified in those of Escherichia coli, Staphylococcus aureus and Y. enteroeolitica (0:9). The detection results indicated that the sensitivity of milk samples reached 50 cfu/mL, and that of purified DNA of Bmcella reached 1.5pg and the repeatability and stability of them were quite higher. Therefore, PCR detection method for pathogen of brucellosis were found highly specific and sensitive and could be used for rapid diagnosis of brucellosis.
机构地区 广西兽医研究所
出处 《广西农业科学》 CSCD 2009年第7期915-918,共4页 Guangxi Agricultural Sciences
基金 广西科技攻关项目(0537008-3C)
关键词 布鲁氏菌病 病原检测 OMP-2基因 PCR brucellosis pathogen detection OMP-2 gene PCR
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