疟原虫新型恒温扩增快速筛检方法的研究

    符丽媛1，宋凌浩1，孟家祺1，朱玲玲1，陆永昌2，陈瑞2，石坚3，胡林3

    1.苏州出入境检验检疫局卫检处，江苏 苏州215021；2.江苏出入境检验检疫局；

    3.杭州优思达生物技术有限公司

    摘要：目的建立可现场使用的疟原虫快速检测方法，为检验检疫基层实验室开展筛检和监控工作提供技术手段。方法通过合成特异引物和探针，系统筛选，优化恒温扩增条件，建立疟原虫核酸等温扩增方法，用免疫层析试纸条快速检测扩增产物，并测试其灵敏度和特异性。结果疟原虫交叉引物恒温扩增方法灵敏度达到103拷贝/ml，反应特异，操作简便，不需要特殊设备，3 h可报告结果。结论本研究应用交叉引物恒温扩增技术（CPA）对疟原虫的核酸进行扩增，并应用试纸条检测技术检测核酸扩增产物，检测时间短，具有一定的应用价值。

    关键词：疟原虫；快速检测；交叉引物恒温扩增技术；研究

    中图分类号：R123.5　文献标识码：B

    Study on rapid detection of plasmodium bycross priming

    amplification(CPA)

    FU Li-yuan*, SONG Ling-hao, MENG Jia-qi, ZHULing-ling, Lu Yong-chang, CHEN Rui, SHI Jian, HU Lin

    *Suzhou Entry-Exit Inspection and QuarantineBureau, Suzhou, Jiangsu 215021,China

    Abstract:   Objective  To establish a method for rapid detection of plasmodium nucleicacid，and to provide suitable method for the inspection andquarantine basic laboratory to carry out plasmodium screening testat entry ports.  Methods   Synthesis of specificprimers and probe, selection of reaction system, optimizingconditions of isothermal amplification were used in theestablishment of rapid detection of plasmodium nucleic acid, andsensitivity and specificity were tested.  Results  The new method was characterized by sensitivity（103copies/ml）,specificity, simple operation, no special equipment required, andthe detection result was reported within three hours.  Conclusion   A new method was developed to amplifyplasmodium nucleic acid by CPA and detect nucleic acidamplification products by the strip. New method can be applied indetecting plasmodium.

    Key words:   Plasmodium; Rapiddetection; Cross priming amplification; Study

    《中国国境卫生检疫杂志》