›› 2001, Vol. 21 ›› Issue (2): 67-69.

• 论文 •    下一篇

大鼠泰泽氏菌套式PCR方法的建立和初步应用

  

  1. 上海市实验动物质量监督检验站 200032
  • 收稿日期:2001-01-03 出版日期:2001-01-25 发布日期:2013-03-19
  • 基金资助:

    上海市科技发展基金资助项目(994919037)

Establishment and Preliminary Application of Nested Polymerase Chain Reaction for Bacillus piliformis

  1. Shanghai Quality Monitoring Center For Laboratory Animals, Shanghai 200032 ,China
  • Received:2001-01-03 Online:2001-01-25 Published:2013-03-19

摘要: 根据秦泽氏菌rDNA序列设计出二对特异引物,以标准菌RJ株和大肠杆菌、溶血性链球菌、嗜肺巴氏杆菌纯化的DNA为模板,进行套式PCR反应。结果用外引物扩增,泰泽氏菌和大肠杆菌均出现625 bp的反应带,其它细菌无反应带;而用内引物第二次扩增,只有泰泽氏菌出现196 bp的特异扩增带。将PCR方法应用于人工免疫抑制的SD大鼠,检出率为30%(9/30),同时将此30份血清用间接免疫荧光法(IFA)进行检测,结果未检出阳性样品。结果提示,套式PCR方法具有特异性强、敏感性高,简便快速的特点。

关键词: 泰泽氏茵, 套式PCR, 免疫抑制, 大鼠, IFA

Abstract: On the basis of the 16S ribosomal DNA(rDNA) sequences, the DNA extract-ed from Bacillus piliformis,Escherichia coli,HemoLytic streptococcus,Pasteuralla pneumotroopica were amplified by PCR using 2 pairs of PCR specific primers. With out-er-primers,a DNA fragment of the expected size (625bp) was amplified from Bacillus piliformis and E.coli, but no band was observed in Hemolytic streptococcus and Pasteu-ralla pyieumotroopica. Whereas a 196bp fragment was obtained only from Bacillus pili-formis with inner-primers. Thirty liver samples from immunosuppressive rats were de-tected for Bacillus piliformis by using nested PCR, the positive rate is 30% (9/30),but no serum sample is positive monitored by IFA. In conclusion, the 16S rDNA-based PCR established is sensitive and specific in detection of Bacillus piliformis in infected ani-mals.

Key words: Bacillus piliformis , Nested PCR , Immunosuppressive , Rat , IFA