建立检测疟疾的微滴数字PCR方法

Development of droplet digital PCR method for malaria detection

目的建立适合口岸疟疾检测的微滴数字PCR方法。方法根据文献中检测疟原虫的特异性引物和探针,建立检测疟疾的微滴数字PCR方法,与普通PCR及荧光定量PCR进行比较,评估微滴数字PCR的优越性。结果微滴数字PCR检测下限为0.721ng/μl,灵敏度比普通PCR高10倍,与荧光定量PCR相当;微滴数字PCR的重复性高于普通PCR,但不如荧光定量PCR稳定。检测的准确性方面,普通PCR对37份已知样本的检测准确率为75.68%,微滴数字PCR和荧光定量PCR均能达到100.00%。结论建立了检测疟疾的微滴数字PCR方法,比普通PCR方法灵敏度、重复性及准确性均有很大优势。

Objective To establish a droplet digital PCR(ddPCR)method for malaria detection at ports.Methods Based on the specific primers and probes for Plasmodium detection in the reference,a ddPCR method for malaria detection was established and compared with ordinary PCR and fluorescent quantitative PCR(qPCR)to evaluate the superiority of ddPCR.Results The detection limit of ddPCR was 0.721 ng/μl,which was the same with q PCR,but was 10 times sensitive than ordinary PCR.The detection repeatability of ddPCR was superior than ordinary PCR,but was not as stable as qPCR.The detection accuracy for 37 identified samples of ordinary PCR was 75.68%,ddPCR and qPCR were all 100.00%.Conclusion A ddPCR method for malaria detection had been established,which has great advantages than ordinary PCR in the sensitivity,repeatability and accuracy.