Serological detection of Plasmodium vivax malaria using recombinant proteins corresponding to the 19-kDa C-terminal region of the merozoite surface protein-1 -: art. no. 39
Serological detection of Plasmodium vivax malaria using recombinant proteins corresponding to the 19-kDa C-terminal region of the merozoite surface protein-1 -: art. no. 39
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DOI：
10.1186/1475-2875-2-39
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发表时间：
2003-11-01
期刊：
影响因子：
3
通讯作者：
Soares, IS
中科院分区：
文献类型：
--
作者：
Rodrigues, MHC;Cunha, MG;Soares, IS
背景资料：检测间日疟原虫特异性抗体的血清学试验可能是流行病学研究、在疟疾非流行地区筛查献血者和诊断受感染个体的宝贵工具。由于间日疟原虫不易于在体外获得，因此很少使用使用全或半纯化抗原的ELISA测定。基于这一局限性，我们测试了代表间日疟原虫裂殖子表面蛋白-1（MSP 1（19））的19 kDa C-末端区域的重组蛋白是否可用于疟疾感染的血清学检测。大肠杆菌中产生的三种纯化的重组蛋白比较了一种在Pichia pastoris中的抗体（GST-MSP 1（19）、His（6）-MSP 1（19）和His（6）-MSP 1（19）-PADRE）和一种在Pichia pastoris中的抗体（yMSP 1（19）-PADRE）与患有间日疟原虫感染的个体的IgG抗体结合的能力。该方法进行了测试，从生活在巴西北部的疟疾流行地区的个人收集的200份血清样本，53个暴露于恶性疟原虫感染的个人的血清样本和177个从未接触到maligna.Results的个人的血清样本：总体而言，ELISA评估的灵敏度与自然感染的个人的血清为95%。与重组蛋白GST-MSP 1（19）、His（6）-MSP 1（19）、His（6）-MSP 1（19）-PADRE和yMSP 1（19）-PADRE反应的血清样品比例分别为90%、93.5%、93.5%和93.5%。用健康人和其他传染病患者血清检测，特异性为98.3%（GST-MSP 1（19））、97.7%（His（6）-MSP 1（19）和His（6）-MSP 1（19）-PADRE）或100%（yMSP 1（19）-PADRE）。我们的研究表明，对于巴西人来说，使用MSP 1（19）的重组蛋白的ELISA可用作开发用于检测间日疟原虫疟疾的有价值的血清学测定的基础。
Background: Serological tests to detect antibodies specific to Plasmodium vivax could be a valuable tool for epidemiological studies, for screening blood donors in areas where the malaria is not endemic and for diagnosis of infected individuals. Because P. vivax cannot be easily obtained in vitro, ELISA assays using total or semi-purified antigens are rarely used. Based on this limitation, we tested whether recombinant proteins representing the 19 kDa C-terminal region of the merozoite surface protein-1 of P. vivax (MSP1(19)) could be useful for serological detection of malaria infection.Methods: Three purified recombinant proteins produced in Escherichia coli (GST-MSP1(19), His(6)-MSP1(19) and His(6)-MSP1(19)-PADRE) and one in Pichia pastoris (yMSP1(19)-PADRE) were compared for their ability to bind to IgG antibodies of individuals with patent P. vivax infection. The method was tested with 200 serum samples collected from individuals living in the north of Brazil in areas endemic for malaria, 53 serum samples from individuals exposed to Plasmodium falciparum infection and 177 serum samples from individuals never exposed to malaria.Results: Overall, the sensitivity of the ELISA assessed with sera from naturally infected individuals was 95%. The proportion of serum samples that reacted with recombinant proteins GST-MSP1(19), His(6)-MSP1(19), His(6)-MSP1(19)-PADRE and yMSP1(19)-PADRE was 90%, 93.5%, 93.5% and 93.5%, respectively. The specificity values of the ELISA determined with sera from healthy individuals and from individuals with other infectious diseases were 98.3% (GST-MSP1(19)), 97.7% (His(6)-MSP1(19) and His(6)-MSP1(19)-PADRE) or 100% (yMSP1(19)-PADRE).Conclusions: Our study demonstrated that for the Brazilian population, an ELISA using a recombinant protein of the MSP1(19) can be used as the basis for the development of a valuable serological assay for the detection of P. vivax malaria.