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猪支气管败血波氏杆菌PCR诊断方法的建立及应用
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杨咏洁，鲁承, 赵达卓，金美伶
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（延边大学 农学院 动物医学系，吉林 龙井133400）
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摘要：
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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237bp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112.5pg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。 |
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关键词：
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猪；支气管败血波氏杆菌； 猪传染性萎缩性鼻炎； 聚合酶链式反应 |
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中图分类号：S 852.614 文献标识码：
A 文章编号：1673-4696(2006)01-0029-04 |
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Development and application of PCR assay for diagnosis of
Bordetella bronchiseptica in pigs |
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YANG Yongjie, LU Cheng, ZHAO Dazhuo, JIN Meiling |
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(Department of Veterinary Medicine, College of Agriculture, Yanbian University, Longjing 133400, China） |
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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 112.5pg 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 microagglutination 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 microagglutination test. |
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Key words: swine; Bordetella bronchiseptica; atrophic rhinitis; PCR |
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