Mirahmadi Hadi, Fallahi Shirzad, Seyyed Tabaei Seyyed Javad
Infectious Diseases and Tropical Medicine Research Center, Zahedan University of Medical Sciences, Zahedan, Iran; Department of Parasitology and Mycology, Faculty of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran.
Razi Herbal Medicines Research Center, Lorestan University of Medical Sciences, Khorramabad, Iran; Department of Parasitology and Mycology, Facultyof Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran.
J Microbiol Methods. 2016 Apr;123:44-50. doi: 10.1016/j.mimet.2016.02.003. Epub 2016 Feb 3.
Enzyme Linked Immunosorbent Assay (ELISA), as a serological test, can be a beneficial tool for epidemiological studies by screening blood donors and diagnosis of specific antibodies from Plasmodium vivax (P. vivax) infected cases. Since P. vivax cannot easily be acquired in vitro, ELISA assays using total or semi-purified antigens are seldom used. On the basis of this restriction, we examined whether recombinant protein 42 kDa related to C-terminal region of the merozoite surface antigen-1 of P. vivax (MSA-1(42)) could be suitable for serological detection of vivax malaria infection. Purified recombinant protein produced in Escherichia coli (E. coli) (GST-MSA-1(42)) was examined for its ability to bind to IgG antibodies of individuals with patent P. vivax infection. The method was tested with 262 serum samples collected from individuals living in the south and southeastern regions of Iran where malaria is endemic. Samples exposed to Plasmodium falciparum (P. falciparum) infection and patients with other infectious disease (toxoplasmosis, Leishmania infantum infection, echinococcosis and FUO (fever with unknown origin)) except for P. falciparum were residing in non- malaria endemic areas in Iran. Generally, the sensitivity of ELISA evaluated with sera from naturally infected individuals was 86.9%. The specificity value of the ELISA determined with sera from healthy individuals and from individuals with other infectious diseases was 94.05%. The positive predictive value (PPV), negative predictive value (NPV) provided, and the diagnostic efficiency of anti-rPvMSA-1(42) antibody using indirect ELISA were determined 93.58, 87.77 and 91.06% respectively. Our study demonstrated that, because MSA-1(42) kDa contains both the 33 and 19 kDa fragments in its structure, it can serve as the basis for the development of a sensitive serological test which can be used for epidemiological studies, screening blood donors and diagnosis of P. vivax malaria.
酶联免疫吸附测定(ELISA)作为一种血清学检测方法,通过筛查献血者以及诊断间日疟原虫(P. vivax)感染病例中的特异性抗体,可成为流行病学研究的有益工具。由于间日疟原虫难以在体外轻易获取,因此很少使用使用全抗原或半纯化抗原的ELISA检测方法。基于这一限制,我们研究了与间日疟原虫裂殖子表面抗原-1(MSA-1)C端区域相关的重组蛋白42 kDa(MSA-1(42))是否适合用于间日疟感染的血清学检测。检测了在大肠杆菌(E. coli)中产生的纯化重组蛋白(GST-MSA-1(42))与间日疟现症感染个体IgG抗体结合的能力。该方法用从伊朗南部和东南部疟疾流行地区居民收集的262份血清样本进行了测试。暴露于恶性疟原虫(P. falciparum)感染的样本以及除恶性疟原虫外患有其他传染病(弓形虫病、婴儿利什曼原虫感染、棘球蚴病和不明原因发热(FUO))的患者来自伊朗非疟疾流行地区。总体而言,用自然感染个体的血清评估的ELISA敏感性为86.9%。用健康个体和患有其他传染病个体的血清测定的ELISA特异性值为94.05%。使用间接ELISA测定的抗rPvMSA-1(42)抗体的阳性预测值(PPV)、阴性预测值(NPV)以及诊断效率分别为93.58%、87.77%和91.06%。我们的研究表明,由于MSA-1(42) kDa在其结构中同时包含33 kDa和19 kDa片段,它可为开发一种敏感的血清学检测方法奠定基础,该方法可用于流行病学研究、筛查献血者以及诊断间日疟。