| 牛瑟氏泰勒虫环介导等温扩增检测方法的建立 |
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| 引用本文： | 王中光,张守发,曹世诺,贾立军,薛书江,田万年,王暖成.牛瑟氏泰勒虫环介导等温扩增检测方法的建立[J].中国预防兽医学报,2010,32(2). |
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| 作者姓名： | 王中光 张守发 曹世诺 贾立军 薛书江 田万年 王暖成 |
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| 作者单位： | 延边大学农学院,动物医学系,吉林,龙井,133400 |
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| 摘 要： | 为建立快捷、灵敏的牛瑟氏泰勒虫(T.sergenti)检测方法,本研究以感染牛T.sergenti的阳性血液为试验材料,建立了牛T.sergenti P33表面蛋白基因环介导等温扩增(LAMP)检测技术,并优化了LAMP的各反应条件,进行了敏感性和特异性试验。结果显示,扩增产物经显色、电泳、酶切鉴定后,LAMP方法扩增牛T.sergenti产物检测管显色后呈阳性反应,最低检测量为2.3fg/μL模板DNA,比普通PCR高10倍,牛T.sergenti LAMP产物电泳后呈特征性梯状条带,而弓形虫等对照组均无扩增条带。说明本研究建立的LAMP方法具有灵敏、特异、快速等优点,适合于牛T.sergenti的检测。
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| 关 键 词： | 牛瑟氏泰勒虫 P33表面蛋白基因 环介导等温扩增技术 |
Development of LAMP for detection of Theileria sergenti |
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| WANG Zhong-guang,ZHANG Shou-fa,CAO Shi-nuo,JIA Li-jun,XUE Shu-jiang,TIAN Wan-nian,WANG Nuan-cheng.Development of LAMP for detection of Theileria sergenti[J].Chinese Journal of Preventive Veterinary Medicine,2010,32(2). |
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| Authors: | WANG Zhong-guang ZHANG Shou-fa CAO Shi-nuo JIA Li-jun XUE Shu-jiang TIAN Wan-nian WANG Nuan-cheng |
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| Institution: | WANG Zhong-guang,ZHANG Shou-fa,CAO Shi-nuo,JIA Li-jun,XUE Shu-jiang,TIAN Wan-nian,WANG Nuan-cheng (Department of Veterinary Medicine,Yanbian University,Longjing 133400,China) |
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| Abstract: | To establish a sensitive and rapid detection method for Theileria sergenti, a loop-mediated isothermal amplification (LAMP) was established for detection of T. sergenti P33 surface protein gene from positive blood samples. LAMP was conducted under optimized conditions and results were visualized by naked eye, electrophoretic analysis and endonuclease digestion. This method with a detection limitation of 2.3 fg/μL DNA is fast, sensitive and specific for detection of T. sergenti. |
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| Keywords: | Theileria sergenti p33 surface protein gene loop-mediated isothermal amplification LAMP |
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