【摘要】 根据支气管败血波氏杆菌(Bordetella bronchiseptica)鞭毛蛋白基因的上游序列设计了1对引物fla1和fla2,扩增出大小为237 bp的目的基因片段,建立了快速检测支气管败血波氏杆菌的PCR方法。特异性和敏感性试验表明,该方法对大肠埃希氏菌、金黄色葡萄球菌和D型多杀性巴氏杆菌均无交叉性反应;最低能够检出112.5 pg的模板DNA。用该PCR法检测了从延边州采集的48份表现不同临床症状的猪鼻黏液;结果,检出支气管败血波氏杆菌阳性42例,阳性率为87.5%。同时与传统的细菌检查法、微量凝集法进行了比较;结果显示,该PCR法的检出率是细菌检查法的5.25倍,是微量凝集法的1.75倍。更多还原

【Abstract】 With a pair of primers designed to amplify flagellin gene of Bordetella bronchiseptica according to the upstream sequence of the gene,a fragment of 237 bp in length of the target gene was amplified by PCR and a standard PCR assay was developed for quick detection of B.bronchiseptica infection.Specificity and sensitivity assays revealed that the assay did not cross react with Escherichia coli,Staphylococcus aureus and Pasteurella multocida type D,and the assay could detect as low as 112.5pg template DNA.48 samples of nasal mucus from pigs from Yanbian Prefecture with different clinical symptoms was examined by the developed PCR assay and 42 samples(87.5%) were positive.Comparison of the assay with the conventional bacterioscopy and the micro-agglutination test revealed that the sensitivity of the PCR assay was 5.25 times higher than that of the bacterioscopy and 1.75 times higher than that of the micro-agglutination test.更多还原