A dot-blot ELISA preliminary evaluation using PvMSP1-42 recombinant protein as antigen for serological diagnosis of Plasmodium vivax infection in Thailand.

European journal of microbiology & immunology Pub Date : 2024-03-01 Print Date: 2024-05-14 DOI:10.1556/1886.2024.00008
Kantima Choosang, Siriphan Boonsilp, Kanyanan Kritsiriwuthinan, Palin Chumchuang, Nanthawan Thanacharoensakun, Aminoh Saai, Sawanya Pongparit
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Abstract

Plasmodium vivax is the most prevalent cause of malaria in Thailand and is predominant in malarial endemic areas worldwide. P. vivax infection is characterized by low parasitemia, latent liver-stage parasites, or asymptomatic infections leading to underreported P. vivax cases. These are significant challenges for controlling and eliminating P. vivax from endemic countries. This study developed and evaluated a dot-blot enzyme-linked immunosorbent assay (ELISA) using PvMSP1-42 recombinant antigen for serological diagnosis based on the detection of antibodies against P. vivax. The optimal PvMSP1-42 concentration and dilutions of anti-human IgG horseradish peroxidase (HRP)-conjugated antiserum were tested on 88 serum samples from P. vivax, Plasmodium falciparum and bacterial infection, including healthy individuals. A cut-off titer of 1:800 produced optimal values for sensitivity and specificity of 90.9 and 98.2%, respectively, with an accuracy of 95.5%. The positive and negative predictive values were 96.8 and 94.7% respectively. The results from microscopic examination and dot-blot ELISA showed strong agreement with the 0.902 kappa index. Thus, the dot-blot ELISA using PvMSP1-42 antigen provided high sensitivity and specificity suitable for serodiagnosis of P. vivax infection. The test is a simple and quick diagnostic assay suitable for field testing as it does not require specific equipment or particular skills.

使用 PvMSP1-42 重组蛋白作为抗原对泰国间日疟原虫感染进行血清学诊断的点印迹 ELISA 初步评估。
间日疟原虫是泰国最常见的疟疾病因,也是全球疟疾流行地区的主要病因。间日疟原虫感染的特点是低寄生虫血症、潜伏肝期寄生虫或无症状感染,导致间日疟病例报告不足。这些对控制和消除流行国家的间日疟来说都是重大挑战。本研究利用 PvMSP1-42 重组抗原开发并评估了一种点印迹酶联免疫吸附试验(ELISA),该试验基于对间日疟原虫抗体的检测进行血清学诊断。在包括健康人在内的 88 份来自间日疟原虫、恶性疟原虫和细菌感染的血清样本上测试了抗人 IgG 辣根过氧化物酶(HRP)结合抗血清的最佳 PvMSP1-42 浓度和稀释度。截止滴度为 1:800 时,灵敏度和特异度的最佳值分别为 90.9% 和 98.2%,准确度为 95.5%。阳性和阴性预测值分别为 96.8% 和 94.7%。显微镜检查和点印迹酶联免疫吸附试验的结果显示出很高的一致性,卡帕指数为 0.902。因此,使用 PvMSP1-42 抗原的点印迹酶联免疫吸附试验具有较高的灵敏度和特异性,适合用于间日疟原虫感染的血清诊断。由于不需要特定的设备或技能,该检验是一种简单、快速的诊断方法,适用于现场检验。
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