兔支气管败血波氏杆菌PCR检测方法的建立及应用

Establishment of PCR Assay for Diagnosis of Bordetella bronchiseptica in Rabbits

根据支气管败血波氏杆菌菌毛蛋白基因(fimN)序列设计了一对引物,扩增出大小为648bp的目的基因片段,建立了快速检测兔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、金黄色葡萄球菌和巴氏杆菌均无交叉性反应。用该PCR法检测了从杨凌、咸阳采集的50份表现临床症状的兔呼吸道分泌物检出支气管败血波氏杆菌阳性35例,阳性率为70%。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3.89倍,是微量凝集法的1.94倍。

With a pair of primers designed to amplify fimbrial gene of Bordetella bronchiseptica according to the sequence of the gene, a fragment of 648bp in length on the target gene was amplified by PCR and a standard PCR assay was developed for quick detection of Bordetella bronchiseptica infection. Specificity and sensitivity assays revealed that the assay cross reaction with Escherichia coli, Staphylococcus and Pasteurella. 50 samples of nasal mucus from rabbits from Yangling and Xianyang were examined by the developed PCR assay and 70% were positive. Comparison of the assay with the conventional bacterioscopy and the micro-agglutination test revealed that the sensitivity of the PCR assay was 3.89 times higher than that of the bacterioscopy and 1.94 times higher than that of the micro-agglutination test