首页> 美国卫生研究院文献>Journal of Clinical Microbiology >Serodiagnosis of Human Granulocytic Ehrlichiosis by a Recombinant HGE-44-Based Enzyme-Linked Immunosorbent Assay
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Serodiagnosis of Human Granulocytic Ehrlichiosis by a Recombinant HGE-44-Based Enzyme-Linked Immunosorbent Assay

机译:基于重组HGE-44的酶联免疫吸附试验对人粒细胞性尿病的血清学诊断

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摘要

Current antibody testing for human granulocytic ehrlichiosis relies predominantly on indirect fluorescent-antibody assays and immunoblot analysis. Shortcomings of these techniques include high cost and variability of test results associated with the use of different strains of antigens derived from either horses or cultured HL-60 cells. We used recombinant protein HGE-44, expressed and purified as a maltose-binding protein (MBP) fusion peptide, as an antigen in a polyvalent enzyme-linked immunosorbent assay (ELISA). Fifty-five normal serum samples from healthy humans served as a reference to establish cutoff levels. Thirty-three of 38 HGE patient serum samples (87%), previously confirmed by positive whole-cell immunoblotting, reacted positively in the recombinant ELISA. In specificity analyses, serum samples from patients with Lyme disease, syphilis, rheumatoid arthritis, and human monocytic ehrlichiosis (HME) did not react with HGE-44–MBP antigen, except for one sample (specificity, 98%). We conclude that recombinant HGE-44 antigen is a suitable antigen in an ELISA for the laboratory diagnosis and epidemiological study of HGE.
机译:当前用于人类粒细胞性大肠杆菌病的抗体测试主要依赖于间接荧光抗体测定和免疫印迹分析。这些技术的缺点包括与使​​用源自马或培养的HL-60细胞的抗原的不同菌株有关的测试结果的高成本和可变性。我们在多价酶联免疫吸附测定(ELISA)中使用重组蛋白HGE-44(作为麦芽糖结合蛋白(MBP)融合肽表达和纯化)作为抗原。来自健康人的55份正常血清样品用作确定临界水平的参考。先前已通过全细胞免疫印迹阳性确认的38个HGE患者血清样品中的33个(87%)在重组ELISA中反应阳性。在特异性分析中,来自莱姆病,梅毒,类风湿性关节炎和人单核细胞埃希氏病(HME)的患者的血清样品与HGE-44–MBP抗原没有反应,除了一种样品(特异性为98%)。我们得出的结论是,重组HGE-44抗原是ELISA中适合HGE实验室诊断和流行病学研究的抗原。

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