恶性疟原虫LF-RPA检测方法研究

STUDY ON LF-RPA DETECTION METHOD OF PLASMODIUM FALCIPARUM

本研究根据恶性疟原虫18S rDNA保守区序列设计引物和探针,建立基于重组酶聚合酶扩增(recombinase polymerase amplification,RPA)与侧流层析(lateral flow,LF)相结合的技术检测恶性疟原虫,同时评价该方法的灵敏性和特异性。结果表明,本方法能够在39℃恒温条件下15 min内有效检测出恶性疟原虫DNA,特异性好,与其他传染病病原无交叉反应,灵敏度为6.54×10^(2)copies/反应,显著高于普通胶体金试纸条检测方法。本研究所建立的方法可用于恶性疟原虫的快速检测。

To establish a novel detection technique for Plasmodium falciparum on the combination of recombinase polymerase amplification(RPA)and lateral flow(LF)chromatography,the primer sets and probes were designed according to the conserved region of 18S rDNA in P.falciparum,and the detection protocol were organized following the assessment of sensitivity and specificity.As results,the newly developed protocol detect P.falciparum DNA effectively within 15 min at 39℃,which indicates the protocol with good specificity and sensitivity,the lower detection limit is near to 6.54×10^(2)copies/reaction,much sensitive than commercial colloidal gold test strip method.The protocol established should be applicable in the rapid and accuracy detection of P.falciparum in clinical and related samples.