疟原虫新型恒温扩增快速筛检方法的研究

Study on rapid detection of plasmodium by cross priming amplification(CPA)

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,3h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。

Objective To establish a method for rapid detection of plasmodium nucleic acid,and to provide suitable method for the inspection and quarantine basic laboratory to carry out plasmodium screening test at entry ports.Methods Synthesis of specific primers and probe,selection of reaction system,optimizing conditions of isothermal amplification were used in the establishment of rapid detection of plasmodium nucleic acid,and sensitivity and specificity were tested.Results The new method was characterized by sensitivity（103copies/ml）,specificity,simple operation,no special equipment required,and the detection result was reported within three hours.Conclusion A new method was developed to amplify plasmodium nucleic acid by CPA and detect nucleic acid amplification products by the strip.New method can be applied in detecting plasmodium.