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当前位置: 首页 > 文章 > 鸭疫里默氏杆菌16 S rRNA PCR快速检测方法的建立 中国兽医科学 2010 (4) 395-399
Position: Home > Articles > Development of PCR based on 16 S rRNA gene for rapid detection of Riemerella anatipestifer Chinese Veterinary Science 2010 (4) 395-399

鸭疫里默氏杆菌16 S rRNA PCR快速检测方法的建立

作  者：
冯金牛;陈芳艳;阮二垒;杨丽云;黎丁滔;陈瑞爱;区德庆;唐秀英;王林川
单  位：
华南农业大学兽医学院;华南农业大学动物科学学院;广东大华农动物保健品股份有限公司
关键词：
鸭疫里默氏杆菌;16 S rRNA基因;聚合酶链反应
摘  要：
根据GenBank中的鸭疫里默氏杆菌(RA)1~19型16 S rRNA基因序列,分别与大肠杆菌、沙门氏菌、葡萄球菌、巴氏杆菌的16 S rRNA基因序列对比,设计特异性引物,建立了基于鸭疫里默氏杆菌16 SrRNA的PCR检测方法。结果表明,用该方法对鸭疫里默氏杆菌、大肠杆菌、沙门氏菌、葡萄球菌、巴氏杆菌进行检测,只有鸭疫里默氏杆菌的DNA能扩增出680 bp的特异条带,其他细菌的DNA不能扩增出任何条带;测序结果与GenBank中鸭疫里默氏杆菌相应序列完全一致;同时分别将鸭疫里默氏杆菌基因组DNA和菌液10倍稀释进行PCR扩增,对鸭疫里默氏杆菌DNA的最小检出量为1.907×10-5g/L,对鸭疫里默氏杆菌菌液的最小检出量为1.5×104CFU/mL。证实,建立的基于鸭疫里默氏杆菌16 S rRNA的PCR检测方法具有快速、灵敏、特异的优点,可用于鸭疫里默氏杆菌感染的检测、分子流行病学调查和分离菌株的快速鉴定。
译  名：
Development of PCR based on 16 S rRNA gene for rapid detection of Riemerella anatipestifer
作  者：
FENG Jin-niu1,CHEN Fang-yan3,RUAN Er-lei1,YANG Li-yun1,LI Ding-tao1,CHEN Rui-ai2,OU De-qing2,TANG Xiu-ying2,WANG Lin-chuan1,2(1.College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China;2.Guangdong Dahuanong Animal Health Products Co.,Ltd.,Xinxing 527400,China;3.College of Animal Science,South China Agricultural University,Guangzhou 510642,China)
关键词：
Riemerella anatipestifer;16S rRNA gene;PCR
摘  要：
Through comparison of 16S rRNA gene sequence of Riemerella anatipestifer serotype 1 to 19 with that of Escherichia coli,Salmonella,Pasteurella and Staphylococci,a pair of specific primers was designed.With the primers,a PCR assay was developed for the detection of R.anatipestifer.The R.anatipestifer,E.coli,Salmonella,Pasteurella and Staphylococci isolates were tested by the developed PCR.A fragment of 680bp was only detected in the R.anatipestifer isolate but no any bands were amplified from the other isolates.The sequences of the 680bp-fragment from the R.anatipestifer isolates were in accordance with that reported in GenBank.Sensitivity was detected by testing 10-fold-diluted R.anatipestifer genomic DNA and cultural liquid,respectively.The detection limit of the R.anatipestifer genomic DNA was 1.907×10-5g/L and the detection limit of the cultural liquid was 1.5×104CFU/mL.The results showed that the developed PCR assay had advantages of rapidity,specificity and sensitivity,and could be used for the detection of R.anatipestifer infection,for the investigation of molecular epidemiology and for the rapid identification of R.anatipestifer isolates.

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