| 简单异尖线虫环介导等温扩增(LAMP)鉴定方法的建立和应用 |
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| 引用本文： | 郑洋妹,陈信忠,龚艳清,杨俊萍. 简单异尖线虫环介导等温扩增(LAMP)鉴定方法的建立和应用[J]. 国际医学寄生虫病杂志, 2009, 37(5): 141-144. DOI: 10.3760/cma.j.issn.1673-4122.2010.03.004 |
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| 作者姓名： | 郑洋妹 陈信忠 龚艳清 杨俊萍 |
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| 作者单位： | 福建农林大学动物科学学院,福州,350002;厦门出入境检验检疫局,厦门,361026; |
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| 基金项目： | 厦门出入境检验检疫局科研基金 |
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| 摘 要： | Objective To establish a simple and quick method of loop-mediated isothermal amplification (LAMP) for identification of Anisakis simplex. Methods According to the ITS gene sequence of A. simplex , a set of specific primers was designed to amplify the specific DNA sequence by LAMP. The concentrations of Mg2+ , Betaine, dNTPs and primers were optimized. The sensitivity and specificity of the method were also tested. Then 7 samples of A. simplex were validated by LAMP. Results The optimal reaction system contained 0. 8 mmol/L dNTPs, 10 mmol/L Mg2+ , 0. 8 mmol/L Betaine, 0. 2 (μmol/L exogenous primers, 1.6 μmol/L inner primers and 8 U Bst DNA polymerase. The optimal reaction temperature was 62 ℃ and reaction time was 60 min. Under those conditions, the detection limit for A. simplex DNA template was 10 copies/μl and no cross reaction with A. typica, Contracaecum sp. , Raphidascaris trichiuri was found. All of the samples of A. simplex were positive by LAMP. Conclusion The LAMP is a sensitive, specific, economic and effective method to identify A. simplex.
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| 关 键 词： | 简单异尖线虫 环介导等温扩增技术 鉴定 |
The establishment of loop-mediated isothermal amplification (LAMP) for identification of Anisakis simplex |
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| ZHENG Yang-mei,CHEN Xin-zhong,GONG Yan-qing,YANG Jun-ping. The establishment of loop-mediated isothermal amplification (LAMP) for identification of Anisakis simplex[J]. International JOurnal of Medical Parasitic Diseases, 2009, 37(5): 141-144. DOI: 10.3760/cma.j.issn.1673-4122.2010.03.004 |
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| Authors: | ZHENG Yang-mei CHEN Xin-zhong GONG Yan-qing YANG Jun-ping |
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| Abstract: | |
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| Keywords: | Anisakis simplexLoop-mediated isothermal amplificationIdentification |
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