| 鸭疫里默氏杆菌16 S rRNA PCR快速检测方法的建立 |
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| 引用本文： | 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.鸭疫里默氏杆菌16 S rRNA PCR快速检测方法的建立[J].中国兽医科学,2010,40(4). |
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| 作者姓名： | 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 |
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| 作者单位： | 冯金牛,阮二垒,杨丽云,黎丁滔,FENG Jin-niu,RUAN Er-lei,YANG Li-yun,LI Ding-tao(华南农业大学兽医学院,广东广州,510642);陈芳艳,CHEN Fang-yan(华南农业大学动物科学学院,广东广州,510642);陈瑞爱,区德庆,唐秀英,CHEN Rui-ai,OU De-qing,TANG Xiu-ying(广东大华农动物保健品股份有限公司,广东新兴,527400);王林川,WANG Lin-chuan(华南农业大学兽医学院,广东广州,510642;广东大华农动物保健品股份有限公司,广东新兴,527400) |
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| 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~(-5)g/L,对鸭疫里默氏杆菌菌液的最小检出量为1.5×10~4CFU/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.
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| 关 键 词： | 鸭疫里默氏杆菌 16 S rRNA基因 聚合酶链反应 |
Development of PCR based on 16 S rRNA gene for rapid detection of Riemerella anatipestifer |
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| FENG Jin-niu,CHEN Fang-yan,RUAN Er-lei,YANG Li-yun,LI Ding-tao,CHEN Rui-ai,OU De-qing,TANG Xiu-ying,WANG Lin-chuan.Development of PCR based on 16 S rRNA gene for rapid detection of Riemerella anatipestifer[J].Veterinary Science in China,2010,40(4). |
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| Authors: | FENG Jin-niu CHEN Fang-yan RUAN Er-lei YANG Li-yun LI Ding-tao CHEN Rui-ai OU De-qing TANG Xiu-ying WANG Lin-chuan |
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| Institution: | 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 527400,3.College of Animal Science,China) |
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| Abstract: | 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 isola... |
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| Keywords: | Riemerella anatipestifer 16 S rRNA gene PCR |
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