Immunoproteomic Identification and Characterization of Leishmania Membrane Proteins as Non-Invasive Diagnostic Candidates for Clinical Visceral Leishmaniasis.
Immunoproteomic Identification and Characterization of Leishmania Membrane Proteins as Non-Invasive Diagnostic Candidates for Clinical Visceral Leishmaniasis.
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DOI：
10.1038/s41598-018-30546-y
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发表时间：
2018-08-14
影响因子：
4.6
通讯作者：
Ali N
中科院分区：
文献类型：
--
作者：
Ejazi SA;Bhattacharyya A;Choudhury ST;Ghosh S;Sabur A;Pandey K;Das VNR;Das P;Rahaman M;Goswami RP;Ali N
内脏利什曼病（VL）是一种潜在的致命疾病，是由多诺瓦利什曼原虫引起的寄生虫感染的结果。本病的临床诊断试验仍与浸润性组织穿刺或血清学免疫层析有关。免疫蛋白质组学的进步，如二维凝胶电泳、质谱、B细胞表位预测和肽合成，使研究人员能够发现用于疾病诊断的新生物标志物。在这项研究中，我们筛选了几种尿反应性利什曼膜蛋白作为潜在的生物标志物候选人。在免疫blot检测中，三种蛋白51、55和63 kDa对47例VL患者的尿液显示100%的反应性，对18例健康和其他疾病无反应。质谱分析显示，51、55和63 kDa蛋白分别为延伸因子1α (EF1-α)、α-微管蛋白和糖蛋白63。通过生物信息学工具绘制这些蛋白的B细胞反应性表位，并从每个蛋白中合成一个得分最高的表位。采用ELISA法检测三种天然电洗脱蛋白及其相应的合成肽与VL和对照尿样的反应性。三者均表现出良好的反应性，其中EF1-α的诊断效果最好。我们的研究结果表明，使用基于尿液的蛋白质组学方法在VL的非侵入性临床诊断中发现生物标志物。
Visceral leishmaniasis (VL), a potentially fatal disease is an outcome of infection caused by the parasite Leishmania donovani. The clinical diagnostic tests for this disease are still related to invasive tissue aspiration or serological immunochromatography. Advancements in immunoproteomics such as two-dimensional gel electrophoresis, mass spectrometry, B cell epitope prediction, and peptide synthesis have enabled researchers to discover newer biomarkers for disease diagnosis. In this study, we have screened several urine-reactive leishmanial membrane proteins as potential biomarker candidates. In the immunoblot assay, three proteins 51, 55 and 63 kDa showed 100% reactivity to the urine of 47 VL patients and nonreactive to 18 healthy and other diseases. Mass spectrometry revealed the identity of 51, 55 and 63 kDa proteins as elongation factor 1α (EF1-α), α-tubulin, and glycoprotein 63, respectively. B cell reactive epitopes of these proteins were mapped through bioinformatic tools and one epitope from each protein that had the highest score were synthesized. All the three native electroeluted proteins and their corresponding synthetic peptides were tested through ELISA for reactivity with VL and control urine samples. While all three demonstrated good reactivity, the diagnostic performance of EF1-α was the best. Our findings illustrate the use of urine-based proteomic approach for biomarker discovery in non-invasive clinical diagnosis of VL.
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DOI：
10.1016/s0732-8893(02)00410-8
发表时间：
2002-08-01
影响因子：
2.9
作者：
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影响因子：
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作者：
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