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基于血清蛋白质组的筛选和抗原应用,用于日本血吸虫病的诊断。

Serological proteome-oriented screening and application of antigens for the diagnosis of Schistosomiasis japonica.

机构信息

The Anhui Provincial Laboratory of Pathogen Biology, Anhui Medical University, Hefei 230032, China.

出版信息

Acta Trop. 2010 Oct;116(1):1-8. doi: 10.1016/j.actatropica.2010.04.014. Epub 2010 May 6.

Abstract

Schistosomiasis remains a major parasitic disease, with 200 million people infected and 779 million people at risk worldwide. The lack of reliable diagnostic techniques makes this disease difficult to control. In an attempt to discover useful candidates for the diagnosis of schistosomiasis, proteomics in combination with western blotting were employed in this study. This serological proteome assay yielded more than 30 immunodominant spots. Ten of these spots were precisely matched with a homologous two-dimensional electrophoresis (2-DE) gel and successfully identified by LC/MS-MS as corresponding to four different proteins. Of these proteins, SjLAP and SjFBPA were successfully expressed, and their recombinant protein products were further applied in the diagnosis of human Schistosomiasis japonica using ELISA. The ELISA results revealed sensitivities of 98.1% and 87.8% for acute and chronic schistosomiasis with rSjLAP and 100% and 84.7% with rSjFBPA, whereas the assays showed a specificity of 96.7% with both recombinant proteins. After treatment with praziquantel, the titres of the antibodies against both antigens declined significantly (P<0.001). Our data therefore suggest that these antibody-oriented recombinant proteins had a high efficacy for the diagnosis of S. japonica, and 2-DE based screening followed by LC/MS-MS has promising potential in the screening of candidate antigens for the diagnosis of schistosomiasis.

摘要

血吸虫病仍然是一种主要的寄生虫病,全球有 2 亿人感染,7.79 亿人面临风险。由于缺乏可靠的诊断技术,这种疾病难以控制。为了发现有助于血吸虫病诊断的有用候选物,本研究采用蛋白质组学结合 Western blot 技术。该血清蛋白质组分析产生了 30 多个免疫优势斑点。其中 10 个斑点与同源二维电泳(2-DE)凝胶精确匹配,并通过 LC/MS-MS 成功鉴定为对应于 4 种不同蛋白质。其中, SjLAP 和 SjFBPA 成功表达,并进一步应用于 ELISA 检测日本血吸虫病患者。ELISA 结果显示,rSjLAP 对急性和慢性血吸虫病的敏感性分别为 98.1%和 87.8%,rSjFBPA 的敏感性分别为 100%和 84.7%,而两种重组蛋白的特异性均为 96.7%。经吡喹酮治疗后,两种抗原的抗体滴度均显著下降(P<0.001)。因此,这些抗体导向的重组蛋白对日本血吸虫病的诊断具有很高的疗效,基于 2-DE 的筛选结合 LC/MS-MS 具有筛选血吸虫病诊断候选抗原的潜在前景。

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