兔支气管败血波氏杆菌PCR检测方法的建立及应用  

Development and Application of PCR Assay for Detection of Bordetella bronchiseptica in Rabbits

作　　者：王欣[1] 恽时锋[1] 王芳[2] 范志宇[2] 

机构地区：[1]南京军区南京总医院比较医学科,南京210002 [2]江苏省农业科学院,南京210014

出　　处：《中国比较医学杂志》2008年第7期40-45,共6页Chinese Journal of Comparative Medicine

基　　金：中国人民解放军南京军区南京总医院科研基金(项目编号:2005113)

摘　　要：目的建立一种简便、快速、敏感、特异的适用于支气管败血波氏杆菌的PCR检测方法。方法根据兔支气管败血波氏杆菌(Bordetella bronchiseptica)的fim2基因序列设计了一对特异性引物,进行PCR扩增、特异性和敏感性试验,并将其应用于临床样品的检测。结果利用该PCR方法扩增出425 bp的目的基因片段,该产物序列与GeneBank上公布的基因序列同源性为100%。特异性试验表明,该方法对大肠埃希氏菌、多杀性巴氏杆菌、魏氏梭菌和金黄色葡萄球菌均无交叉性反应;并且最小可检出菌液浓度为3.6 CPU。用该PCR方法检测了从江苏、山东等地采集的146份兔鼻拭子,结果检出支气管败血波氏杆菌阳性92例,阳性率为63.01%。结论建立了快速检测支气管败血波氏杆菌的PCR方法。Objective To develope a simple, rapid, sensitive and specific PCR detective method for BordeteUa bronchiseptica. Methods A pair of specific primers was designed to amplify tim2 gene of Bordetella bronchiseptica in rabbits and perform specificity and sensitivity assays. Some clinical samples were used to evaluate the PCR assay. Results A fragment of 425 bp in length of the target gene was amplified by the PCR assay. The sequencing result showed that the homology was 100% compared with the reference sequence published in GeneBank. Specificity assays revealed that the assay did not cress react with Escherichia coli , PasteureUa multocida , Clostridieum welchii and Staphylococcus aureus . The least amount of the bacteria could be detected was 3.6 CPU. 146 samples of nasal mucus from rabbits from Jiangsu and Shandong province were examined by the developed PCR assay.92 samples were positive and the positive rates were 63.01%. Conclusion A PCR assay was developed for quick detection of Bredetella bronchiseptica infection.

关 键 词：兔 支气管败血波氏杆菌 PCR 

分 类 号：S852.614[农业科学—基础兽医学]