疟原虫PCR检测方案改进初探

Modification of PCR method for detection of malaria parasites

目的对目前沿用的检测技术方案予以改进,同时评估新技术方案替代原方案的可行性,探讨2种方案在检出水准及其它因素上的差异。方法采用2种不同实验方案对同一批样本进行检测比对,对原方案(散装试剂方案)的主要技术参数与改进后的新方案(预混试剂方案)进行比对,同时辅以经济学层面的探讨。结果散装试剂方案得分1分,预混试剂方案得分4分。在可视性方面,使用疟原虫种特异性引物对血样进行PCR检测,扩增出预期大小约205bp的恶性疟条带和120bp的间日疟条带,2种试剂方案结果基本一致,均较易辨识。结论采用预混方案占优,在疟疾PCR检测方面有推广前景,并有替代原方案的可行性。

Objective To modify the traditional method of bulk regent for detection of malaria parasites and evaluate thefeasibility of the mixed reagent method to replace the traditional method and discuss the differences in the detection level andother factors between them. Methods Two different experimental methods were used to comparative and detect thehomogeneity of the sample. The main parameters and other factors by two methods were compared and analyzed. Results Thebulk reagents gained 1 score, the mixed reagent scheme gained 4 scores. The PCR results of specific detection demonstratedthat the sample amplified an expected enhanced Plasmodium falciparum band of about 205 bp in size and 120 bp ofPlasmodium vivax band. There was no difference in the PCR results between them. Both were identified easily. Conclusion The new method using mixed reagent is advantageous and it＇s feasible to replace the traditional method.