霍乱弧菌CT毒素环介导等温扩增技术 快速检测方法的建立

吴晓芳 徐德顺 钟芬芳

吴晓芳, 徐德顺, 钟芬芳. 霍乱弧菌CT毒素环介导等温扩增技术 快速检测方法的建立[J]. 疾病监测, 2010, 25(7): 580-584. doi: 10.3784/j.issn.1003-9961.2010.07.023
引用本文: 吴晓芳, 徐德顺, 钟芬芳. 霍乱弧菌CT毒素环介导等温扩增技术 快速检测方法的建立[J]. 疾病监测, 2010, 25(7): 580-584. doi: 10.3784/j.issn.1003-9961.2010.07.023
WU Xiao-fang, XU De-shun, ZHONG Fen-fang. Sensitive and rapid detection of cholera toxin-producing Vibrio cholerae by using loop-mediated isothermal amplification[J]. Disease Surveillance, 2010, 25(7): 580-584. doi: 10.3784/j.issn.1003-9961.2010.07.023
Citation: WU Xiao-fang, XU De-shun, ZHONG Fen-fang. Sensitive and rapid detection of cholera toxin-producing Vibrio cholerae by using loop-mediated isothermal amplification[J]. Disease Surveillance, 2010, 25(7): 580-584. doi: 10.3784/j.issn.1003-9961.2010.07.023

霍乱弧菌CT毒素环介导等温扩增技术 快速检测方法的建立

doi: 10.3784/j.issn.1003-9961.2010.07.023
详细信息
    作者简介:

    吴晓芳,女,浙江省湖州市人,主要从事微生物检验工作

    通讯作者:

    吴晓芳,Tel:0572-2760125,Email:xf980718@126.com

Sensitive and rapid detection of cholera toxin-producing Vibrio cholerae by using loop-mediated isothermal amplification

  • 摘要: 目的 将一种新颖的核酸扩增技术环介导等温扩增技术(LAMP)应用于霍乱弧菌毒素基因ctxA的快速检测。 方法 针对霍乱弧菌毒素基因ctxA设计6条特异性引物(两条内引物、两条外引物和两条环引物)进行LAMP扩增,对扩增反应进行优化。并考核方法的敏感性和特异性。 结果 最佳反应时间为60 min,反应温度为65 ℃。对8种相关肠道细菌进行LAMP扩增,仅含毒素基因ctxA的霍乱弧菌得到阳性扩增结果,证明引物具有很高的特异性。霍乱弧菌基因组DNA和纯培养物的检测灵敏度分别约为68 fg和42 cfu/ml。对模拟样品进行直接检测,检测限为72 cfu/g。 结论 该方法检测霍乱弧菌毒素基因ctxA特异性强、灵敏度高,并且操作简便、检测成本低,1 h即可完成,适合基层检验部门使用。
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出版历程
  • 收稿日期:  2010-01-02
  • 刊出日期:  2010-07-30

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