猪萎缩性鼻炎支气管败血波氏杆菌PCR检测方法的建立

Establishment of PCR method for detection of Bordetella bronchiseptica isolates from swine atrophic rhinitis cases

对表现猪萎缩性鼻炎临床症状的猪群中分离得到的34株支气管败血波氏杆菌(Bordetella bronchiseptica,Bb),采用针对Bb flagellum gene的一对引物进行PCR扩增,结果所有分离物均能扩增出237bp特异性DNA条带,与传统生化鉴定结果相一致,且其最小检出量为0.64pg;而猪鼻腔和肺组织中常见的多杀性巴氏杆菌、金黄色葡萄球菌、枯草芽胞杆菌、铜绿假单胞菌、变形杆菌及大肠埃希氏菌均未能出现任何DNA条带。这表明,本试验建立的PCR方法具有特异性强、灵敏度高、可靠性好等特点,可用于猪萎缩性鼻炎的临床诊断。

Thirty four strains of Bordetella bronchiseptica （Bb） isolated from pig herd showing clinical symptoms of atrophic rhinitis （AR） were identified by PCR using a pair of primer of Bb flagellum gene, A 237 bp specific DNA fragment was detected in all isolates, with a minimum detection sensitivity of 0.64 pg. There was no cross reaction with P.multocida, S.aureus, B.subtilis, P.aeruginosa, P.mirabillis and E.coli, which were commonly found in nasal cavity and lung tissue of pig. These results indicated that this PCR was a specific, sensitive and reproducible method that could be used for clinical diagnosis of AR infection.