Objective To establisha 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 peripheralblood samples from pregnant women were tested .Results The products of LAMP demonstrated the typical ladder patterns when the ？temple was Toxoplasma gondii DNA and digested fragments were found with the predicted sizes .The detection limit reached 10 copies .Time-to-positive and concentration of template had a good linear relationship in the stand curve made by plasmids .Detected with established LAMP method ，two positive samples were found in 57 pregnant women .Conclusion The results indicated that this LAMP system not only had good specificity ，sensitivity ，but simple ，rapid and low-cost ，and might play a role in clinical detection .%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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