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中国农学通报

中国农学通报 ›› 2010, Vol. 26 ›› Issue (1): 9-11.

所属专题： 生物技术

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

菌落聚合酶链反应检测支气管败血波氏杆菌

李伟杰,赵耘,杜昕波,康凯,陈敏,陈永林   

  • 收稿日期:2009-07-28 修回日期:2009-08-22 出版日期:2010-01-05 发布日期:2010-01-05
  • 基金资助:

    国家科技支撑计划

Development of PCR assay for detection of Bordetella bronchiseptica

  • Received:2009-07-28 Revised:2009-08-22 Online:2010-01-05 Published:2010-01-05

PDF (PC)

1066

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1对引物Fla1和Fla2，采用菌落PCR方法扩增目的基因片段来检测支气管败血波氏杆菌。利用该方法成功的从8株支气管败血波氏杆菌中扩增出237 bp左右的特异性目的片段。特异性试验表明，该方法对大肠埃希菌、胸膜肺炎放线杆菌、多杀性巴氏杆菌、猪链球菌和副猪嗜血杆菌均无交叉性反应。灵敏性试验表明，将单个菌落稀释105倍利用此菌落PCR仍能扩增到相应的目的片段。结果表明建立的菌落PCR方法对支气管败血波氏杆菌的检测敏感性高、特异性强，可用于Bb感染的诊断和流行病学调查。

关键词: 木质纤维素, 木质纤维素, 稀酸水解, 金属盐助催化剂, 还原糖

Abstract:

A pair of primers was designed according to the upstream sequence of the flagellin gene of Bordetella bronchiseptica, which specifically amplified a fragment of 237 bp in length of the target gene. Specificity assays revealed that the assay didn’t cross react with Escherichia coli, Actinobacillus pleuropneumoniae, Pasteurella multocida, Streptococcus suis and Haemophilus parasuis. Sensitivity assays showed that the expected sequences were obtained from B. bronchiseptica individual colony diluted to 105 times. The results showed that this PCR technique was specific and sensitive, could apply to Bb diagnoses and epidemiology investigation.