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间日疟原虫裂殖子表面蛋白1作为酶联免疫吸附测定(ELISA)诊断疟疾抗原的效能

Efficacy of the merozoite surface protein 1 of Plasmodium vivax as an antigen for ELISA to diagnose malaria.

作者信息

Kim Yong Man, Hwang Hyun Ah, Yun Woo Sang, Kim Suk-Il, Lee Kil Whoan, Park Seung Kyu, Lee Young Jin, Kim Tae Kyun, Wongsrichanalai Chansuda, Sakanari Judy A, Park Hyun

机构信息

Department of Parasitology, College of Medicine, Wonkwang University, Iksan 570-749, Korea.

出版信息

Yonsei Med J. 2004 Feb 29;45(1):129-34. doi: 10.3349/ymj.2004.45.1.129.

Abstract

Malaria is still a major health problem in Thailand and its incidence is currently rising in Korea. To identify a useful antigen for the diagnosis of malaria patients, a cDNA expression library from malaria parasites was constructed and screened out immunologically. One clone was selected in view of its predominant reactivity with the patient sera. The recombinant malaria parasite antigen (Pv30) with 27 kDa as a C-terminal His-tag fusion protein that was produced in Escherichia coli was identified through immunoblot analysis. The deduced amino acid sequence had the sequence homology with the merozoite surface protein 1 (MSP1) genes of Plasmodium falciparum and P. yoelii, each by 41% and 42%, respectively. Measurement of serum IgG and IgM antibody to Pv30 by enzyme-linked immunosorbent assay (ELISA) was evaluated as a serodiagnostic test for malaria patients in Thailand (endemic area) and Korea (recently reemerging area). The sensitivity of P. vivax, P. falciparum, and P. malariae was 96.3% (26 /27), 90.6% (29/32), and 100% (6/6), respectively, and the specificity was 63.5% (40/63) in Thailand samples. The sensitivity of P. vivax was 98.8% (88/89), and the specificity was 96.6% (86/89) in Korean samples. Pv30 appears to be a good and reliable recombinant antigen for serodiagonosis of malaria in a nonendemic area.

摘要

疟疾仍是泰国的一个主要健康问题,目前在韩国其发病率也在上升。为了鉴定一种用于诊断疟疾患者的有用抗原,构建了疟原虫的cDNA表达文库并通过免疫筛选。鉴于其与患者血清的主要反应性,选择了一个克隆。通过免疫印迹分析鉴定了在大肠杆菌中产生的具有27 kDa的重组疟原虫抗原(Pv30),其为C末端His标签融合蛋白。推导的氨基酸序列与恶性疟原虫和约氏疟原虫的裂殖子表面蛋白1(MSP1)基因具有序列同源性,分别为41%和42%。通过酶联免疫吸附测定(ELISA)检测血清中针对Pv30的IgG和IgM抗体,作为泰国(流行地区)和韩国(最近重新出现疟疾的地区)疟疾患者的血清学诊断试验进行评估。在泰国样本中,间日疟原虫、恶性疟原虫和三日疟原虫的敏感性分别为96.3%(26/27)、90.6%(29/32)和100%(6/6),特异性为63.5%(40/63)。在韩国样本中,间日疟原虫的敏感性为98.8%(88/89),特异性为96.6%(86/89)。Pv30似乎是用于非流行地区疟疾血清学诊断的一种良好且可靠的重组抗原。

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