鸭疫里默氏菌病和大肠杆菌病鉴别诊断双重PCR方法的建立和应用

畜牧兽医学报 ›› 2008, Vol. 39 ›› Issue (4): 517-521.

鸭疫里默氏菌病和大肠杆菌病鉴别诊断双重PCR方法的建立和应用

覃宗华，蔡建平，吕敏娜，余劲术，吴彩艳，温列娜，谢明权

1.广东省农业科学院兽医研究所，广州 510640；2.广东省兽医公共卫生实验室，广州 510640

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2008-04-24 发布日期:2008-04-24

Establishment and Application of Dulex PCR Assay for Differentiating Riemerella anatipestifer and Escherichia coli Infection in Ducks

QIN Zong-hua，CAI Jian-ping，LV Min-na，YU Jin-shu，WU Cai-yan，
WEN Lie-na，XIE Ming-quan

1.Institute of Veterinary Medicine, Guangdong Academy of Agricultural Sciences，Guangzhou 510640，China；2.Guangdong Key Laboratory of Veterinary Public Health,Guangzhou 510640,China

Received:1900-01-01 Revised:1900-01-01 Online:2008-04-24 Published:2008-04-24

摘要/Abstract

摘要： 本研究从鸭疫里默氏菌提取基因组DNA，采用细菌16S rRNA基因的通用引物，用PCR方法成功扩增出鸭疫里默氏菌的部分16S rRNA基因，克隆及测序结果表明其全长1 512 bp，G+C含量为51%，该序列已登入GenBank，登录号为DQ078779。同时利用生物软件对GenBank收录的鸭疫里默氏菌和大肠杆菌的16S rRNA基因序列进行分析后，设计了鸭疫里默氏菌和大肠杆菌的种特异性引物，分别能从各自的16S rRNA基因中扩增出长度为342和718 bp的DNA片段。并据此建立了一种能快速准确鉴别诊断鸭疫里默氏菌和大肠杆菌病的双重PCR方法，该方法能从肝脏组织悬液煮沸后的上清、细菌培养物或提取的细菌基因组DNA中快速扩增出相应的片段，进行鉴别诊断。

关键词: 鸭疫里默氏菌, 大肠杆菌, 16S rRNA, 鉴别诊断

Abstract: In this study, partial sequence of Riemerella anatipestifer 16S rRNA gene was amplified by polymerase chain reaction(PCR) using the bacteria 16S rRNA gene universal primers, which its length was 1 512 basepair (bp) and it has been recorded in GenBank and named as DQ078779. The BLAST analysis indicated that DQ078779 was the most similarity with reported 16S rRNA gene of R. anatipestifer. Moreover, speciesspecific primers were designed according to BLAST results of different 16S rRNA gene sequences of R. anatipestifer and E. coli, which 354 and 718 bp specific DNA fragment could be amplified by the two pairs of primers from R. anatipestifer and E. coli, respectively. Finally, dulex PCR with the specific primer for differentiating R. anatipestifer and E. coli infection in ducks was established, and genomic DNA or cultured bacteria or boiled liver tissue of ducks infected by R. anatipestifer and E. coli could be used as template.

Key words: Riemerella anatipestifer, Escherichia coli, 16S rRNA, differentiating diagnosis

覃宗华;蔡建平;吕敏娜;余劲术;吴彩艳;温列娜;谢明权. 鸭疫里默氏菌病和大肠杆菌病鉴别诊断双重PCR方法的建立和应用[J]. 畜牧兽医学报, 2008, 39(4): 517-521.

QIN Zong-hua;CAI Jian-ping;LV Min-na;YU Jin-shu;WU Cai-yan;WEN Lie-na;XIE Ming-quan. Establishment and Application of Dulex PCR Assay for Differentiating Riemerella anatipestifer and Escherichia coli Infection in Ducks[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2008, 39(4): 517-521.

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