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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Comparison of Two Recombinant Major Outer Membrane Proteins of the Human Granulocytic Ehrlichiosis Agent for Use in an Enzyme-Linked Immunosorbent Assay
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Comparison of Two Recombinant Major Outer Membrane Proteins of the Human Granulocytic Ehrlichiosis Agent for Use in an Enzyme-Linked Immunosorbent Assay

机译：人粒细胞埃希氏病病菌的两种重组主要外膜蛋白用于酶联免疫吸附测定的比较

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Enzyme-linked immunosorbent assay (ELISA) for human granulocytic ehrlichiosis (HGE) using two different recombinant P44 proteins (rP44 and rP44-2hv) of the HGE agent as antigens was evaluated. Sera from a total of 72 healthy humans both from regions where HGE is nonendemic and regions where HGE is endemic were used as negative controls to determine the cutoff value for ELISA. Sera from a total of 14 patients (nine from whom the HGE agent was isolated and five who were HGE-PCR positive) were used as positive controls. One hundred nine sera from 72 patients in an area where HGE is endemic who were suspected of having HGE were examined by ELISA and indirect immunofluorescence assay (IFA). All IFA-negative sera were negative by both ELISAs. Of 39 sera that were IFA positive, 35 and 27 were positive by ELISA using rP44 and rP44-2hv, respectively, indicating that the use of rP44 is more sensitive. Western blot analysis of the four rP44-ELISA-negative IFA-positive sera using whole HGE agent as antigen suggests that these four sera were false IFA positive. There was no difference in results with or without the preabsorption of sera with Escherichia coli or with or without the cleavage of the fused protein derived from the vector. There was a significant positive correlation between IFA titers and optical densities of ELISAs. Four Ehrlichia chaffeensis-positive and 10 Borrelia burgdorferi-positive sera were negative by ELISA. However, twoBabesia microti-positive sera showed strong cross-reactivity to the fused vector protein, which was eliminated after cleavage of the protein. Thus, ELISA using rP44 nonfusion protein would provide a simple, specific, and objective HGE serologic test which can be easily automated.
机译：使用HGE试剂的两种不同重组P44蛋白（rP44和rP44-2hv）作为抗原，评估了人类粒细胞埃希氏菌病（HGE）的酶联免疫吸附测定（ELISA）。来自HGE非流行地区和HGE流行地区的总共72名健康人的血清用作阴性对照，以确定ELISA的临界值。将总共​​14例患者的血清（其中9例从HGE试剂中分离出来，5例HGE-PCR阳性）作为阳性对照。通过ELISA和间接免疫荧光法（IFA）对疑似HGE流行地区HGE流行地区的72名患者的109份血清进行了检查。通过两种ELISA，所有IFA阴性血清均为阴性。在使用rP44和rP44-2hv进行ELISA的39份IFA阳性血清中，分别有35份和27份呈阳性，这表明使用rP44更为敏感。使用完整的HGE试剂作为抗原的四个rP44-ELISA阴性IFA阳性血清的蛋白质印迹分析表明，这四个血清均为假IFA阳性。用或不用大肠杆菌预吸附血清，或不切割源自载体的融合蛋白，结果均无差异。 IFA滴度与ELISA的光密度之间存在显着的正相关。 ELISA法检测到4株恰菲埃里希氏菌阳性和10株伯氏疏螺旋体阳性。但是，两个小巴贝氏菌阳性血清对融合的载体蛋白表现出很强的交叉反应性，该蛋白在切割后被消除。因此，使用rP44非融合蛋白的ELISA将提供简单，特异性和客观的HGE血清学检测，可轻松实现自动化。

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