牛布鲁菌病奶液PCR检测方法的建立及初步应用  

Development and preliminary application of the milk-PCR method to detect Nucleic acid of bovine brucellosis

作　　者：李彦伟[1] 高明春[1] 张文龙[1] 王倩[1] 王君伟[1] K. Nielsen YU Wei-ling 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030 [2]加拿大食品检验局动物疾病研究所,加拿大安大略洲K2H8P9

出　　处：《中国兽医杂志》2011年第4期25-27,共3页Chinese Journal of Veterinary Medicine

基　　金：国家科技支撑计划(2007BAD60B02);现代农业产业奶牛技术体系建设专项资金资助(nycytx-0303);黑龙江省重大科技计划(GA06B202);东北农区奶业集约化生产技术集成与产业示范(2006BAD04A09)

摘　　要：为探讨以牛奶为材料检测牛布鲁菌感染的可行性,本试验建立了可鉴定布鲁菌属的单重PCR以及可鉴定布鲁菌种的多重PCR。结果显示,单重PCR方法的特异性强,只特异性的扩增布鲁菌DNA,对照菌株大肠杆菌、牛败血性波氏杆菌、根瘤农杆菌和苍白杆菌的扩增结果均为阴性;奶液中细菌检测灵敏度为1.08×104CFU/mL。用建立的多重PCR对7株不同种的布鲁菌体和基因组进行鉴定,表明该方法可以区分不同种布鲁菌。本试验中建立的PCR方法能够鉴定所有致病性布鲁菌并对疫苗株和野生毒株加以区分,适用于实验室布鲁菌的快速鉴定。To detect the possibility of Brucella contamination in milk,a single PCR and a multi-PCR were developed.The results showed high specificity,only amplified Brucella.spp.No specific bands could be amplified from control bacteria such as E.coli,Bordetella bronchiseptica,Agrobacterium,Ochrobactrum.The sensitivity of this method was 1.08×104CFU/mL.Seven strains of Brucella could be differentiated by this assay successfully.This study showed that the method can distinguish Brucella.spp.In our study the PCR method can identify all pathogenic strains and distinguish Brucella vaccine and wild strains.This assay provides a rapid way for the identification of brucellosis.

关 键 词：布鲁菌病 奶液 单重PCR 多重PCR 

分 类 号：S855.12[农业科学—临床兽医学]