支气管败血波氏杆菌PCR检测方法的建立和应用  

APPLICATION AND DEVELOPMENT OF PCR ASSAY TO DETECT BORDETELLA BRONIHSEPTICA

作　　者：肖海君[1] 崔金生[1] 翁立雪[1] 王小娥[1] 杨春晓[1] 曲朝杰[1] 朱彩霞[1] 刘冠华[1] 朱瑞良[1] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018

出　　处：《山东农业大学学报（自然科学版）》2009年第1期70-74,共5页Journal of Shandong Agricultural University：Natural Science Edition

基　　金：国家自然基金(No.30670114;No.30740077);山东省自然基金项目(No.Y2006D09)资助

摘　　要：根据支气管败血波氏杆菌ptxA基因设计1对引物,扩增特异的872bp核酸片段,建立了应用PCR检测支气管败血波氏杆菌的方法。特异性试验结果表明,3株支气管败血波氏杆菌参考菌株均能扩增出872bp特异性的核酸片段,兔源性的大肠杆菌、巴氏杆菌、沙门氏菌、金黄色葡萄菌的扩增结果均为阴性。敏感性试验结果表明,PCR的最低检出限量为210个支气管败血波氏杆菌。利用建立的PCR检测方法对22株从山东省泰安地区某兔场采集的疑似兔支气管败血波氏杆菌病病兔鼻黏液棉拭子进行检测,结果19株为阳性。并同时进行普通血清学方法和细菌学方法,对检测结果进行了比较,结果显示,PCR方法阳性检出率高于其他检测方法。The ptxA gene recently discovered was shown to be species - specific. A pair of primers was designed to specifically amplified a 872 bp fragment. The PCR result of specificity assay showed that three references Bordetella bronchisepticas could be detected by the PCR test, But no amplification was observed when other 4 bacterial species isolated from rabbits tissue were detected. And the sensitivity result showed that the minimum dose of Bordetella bronchiseptica that could be detected by PCR assay was 210 bacterias. 22 clinical sample, rom one rabbits farm in Tai＇an of Shandong province, doubtedly infected with Bordetella bronchiseptica were tested by PCR. The results showed that 20 postive samples could be detected by the PCR assay. At the same time, serology test and conventional bacterioscopy were also used for the clinical samples. The study provinced a quickly and accurately comparison of the assay with the conventional bacterioscopy and the serology test revealed that the sensitivity of the PCR assay was higher than that of the bacterioscopy and the serology test.

关 键 词：兔 支气管败血波氏杆菌 PCR 检测 建立 应用 

分 类 号：S852.61[农业科学—基础兽医学]