| 环介导等温扩增检测弓形虫方法的建立 |
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| 引用本文： | 陈道桢,许飞,葛海燕,徐兰兰,潘淑静,朱斌. 环介导等温扩增检测弓形虫方法的建立[J]. 中国实验诊断学, 2011, 15(10): 1712-1715 |
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| 作者姓名： | 陈道桢 许飞 葛海燕 徐兰兰 潘淑静 朱斌 |
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| 作者单位： | 1. 南京医科大学附属无锡妇幼保健院,中心实验室,江苏,无锡214002
2. 苏州大学基础医学与生物科学学院,医学遗传研究室,江苏,苏州215123 |
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| 基金项目： | 江苏省科技厅社会发展资助项目 |
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| 摘 要： | 目的建立环介导等温扩增（LAMP）检测弓形虫DNA的方法。方法以构建的弓形虫B1基因质粒为阳性模板,建立LAMP检测弓形虫的方法,并进行特异性和敏感性评估,在此基础上建立LAMP检测弓形虫的定量体系及通过显色反应直接判断结果的简便方法,最后应用建立的方法对57例孕妇外周血标本进行检测。结果建立的方法仅对弓形虫DNA扩增出特有的梯状条带,酶切结果与预期相符;检测下限为每反应管10拷贝。以重组质粒制作的标准曲线,阳性反应时间与模板浓度有良好的线性关系。57例孕妇标本检测,发现2例阳性。结论本方法特异性强、敏感性高、简便快速且成本低,有望在临床检测中发挥一定的作用。
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| 关 键 词： | 环介导等温扩增 弓形虫 基因诊断 定量检测 |
Establishment of loop-mediated isothermal amplification method for detection of Toxoplasma gondii |
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| CHEN Dao-zhen,XU Fei,GE Hai-yan,et al.. Establishment of loop-mediated isothermal amplification method for detection of Toxoplasma gondii[J]. Chinese Journal of Laboratory Diagnosis, 2011, 15(10): 1712-1715 |
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| Authors: | CHEN Dao-zhen XU Fei GE Hai-yan et al. |
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| Affiliation: | CHEN Dao-zhen1,XU Fei1,GE Hai-yan2,et al.(1.Central Laboratory,Wuxi Hospital for Matemaland Child Health Care Affiliated Medical School of Nanjin,Wuxi,Jiangsu,214002,China,2.Department of preclinical medicine and bioscience,Suchow university,Jiangsu Province,Suchow 215123,China) |
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| Abstract: | Objective To establish a method of Loop-mediated isothermal Amplification(LAMP) for detection of Toxoplasma gondii DNA.Methods Firstly B1 gene plasmids were constructed as positive template,and then the system of LAMP to detect Toxoplasma gondii DNA were established.The specificity and sensitivity were evaluated.A simple method to observe the results and a quantitative protocol were set up.Finally peripheral blood samples from pregnant women were tested.Results The products of LAMP demonstrated the typical ... |
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| Keywords: | LAMP Toxoplasma gondii gene diagnosis quantitative detection |
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