| 牛附红细胞体单管套式PCR检测方法的建立 |
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| 引用本文： | 邢莹,郑秀红,贾立军,薛书江,张守发.牛附红细胞体单管套式PCR检测方法的建立[J].畜牧与兽医,2011,43(4). |
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| 作者姓名： | 邢莹 郑秀红 贾立军 薛书江 张守发 |
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| 作者单位： | 延边大学农学院动物医学系,吉林,延吉,133002 |
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| 基金项目： | 国家自然科学基金项目，公益性行业(农业)科研专项项目 |
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| 摘 要： | 为快速、准确地检测出牛附红细胞体,根据GenBank上发表的牛温氏附红细胞体(Mycoplasma wenyonii)l6S rRNA基因序列(登录号AF016546)设计合成内外2对引物,建立了牛附红细胞体单管套式PCR检测方法,并进行了特异性、敏感性及应用试验。结果:建立的牛附红细胞体单管套式PCR检测方法扩增的片段大小为361 bp,与GenBank中相应序列同源性为98%,该方法扩增不出牛瑟氏泰勒虫、新孢子虫、布氏杆菌及牛无乳链球菌等基因片段,检测出标准模板DNA的最小量为1.22 fgμ/L。通过对吉林省延边地区66份血液样本的临床检测显示,单管套式PCR检出率28.8%,高于常规PCR的21.2%和鲜血压片镜检的12.1%,具有准确、特异、敏感等优点,是检测牛附红细胞体的一种新型、可靠的诊断技术。
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| 关 键 词： | 牛附红细胞体 l6S rRNA 单管套式PCR |
Establishment of a single-tube nested PCR assay for Mycoplasma wenyonii |
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| XING Ying,ZHENG Xiu-hong,JIA Li-jun,XUE Shu-jiang,ZHANG Shou-fa.Establishment of a single-tube nested PCR assay for Mycoplasma wenyonii[J].Animal Husbandry & Veterinary Medicine,2011,43(4). |
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| Authors: | XING Ying ZHENG Xiu-hong JIA Li-jun XUE Shu-jiang ZHANG Shou-fa |
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| Institution: | XING Ying,ZHENG Xiu-hong,JIA Li-jun,XUE Shu-jiang,ZHANG Shou-fa(Department of Veterinary Medicine,Agricultural College of Yanbian University,Yanji 133002,China) |
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| Abstract: | To establish a single-tube nested PCR method for the detection of Mycoplasma wenyonii,two pairs of primers were designed according to the published sequence of the 16S ribosomal RNA gene of M.wenyonii from GenBank(AF016546).Sequencing analysis showed that the targeted gene fragment was 361 bp in length and 98% identical to the published data.The nested PCR method was specific and there were no cross reactions with Theileria sergenti,Neospora caninum,Bacterium burgeri and Streptococcus agalactiae.The sensiti... |
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| Keywords: | 16S rRNA |
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