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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Evaluation of an Immunoglobulin M Capture Enzyme-Linked Immunosorbent Assay for Diagnosis of Orientia tsutsugamushi Infection
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Evaluation of an Immunoglobulin M Capture Enzyme-Linked Immunosorbent Assay for Diagnosis of Orientia tsutsugamushi Infection

机译：免疫球蛋白M捕获酶联免疫吸附试验对虫东方型感染的诊断价值

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摘要

To differentiate scrub typhus from other acute febrile diseases, a rapid and reliable serological diagnosis is important. We developed an immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay (ELISA) for diagnosis of recent Orientia tsutsugamushi infections in humans. The 56-kDa major outer membrane protein of O. tsutsugamushi is well known as the most immunodominant antigen in scrub typhus. The test is based on the use of the biotinylated recombinant 56-kDa protein of O. tsutsugamushi Boryong, Bor56, which was expressed as a fusion protein with a maltose-binding protein in Escherichia coli. In the test, the serum IgM antibodies were captured by anti-human IgM antibodies coated onto a microtiter plate. The captured IgM antibodies were revealed through sequential addition of biotinylated Bor56 antigen and peroxidase-conjugated streptavidin to the plate. The IgM capture ELISA was compared with the immunofluorescence antibody assay (IFA) by testing 176 serum samples from patients with diagnosed cases of rickettsial disease and patients with other acute febrile diseases. Of the 81 IgG IFA-positive samples, 78 tested positive (sensitivity, 96.3%) and all 31 IgM IFA-positive samples tested positive (sensitivity, 100%) by the IgM capture ELISA. The specificity of the IgM capture ELISA was 99%, and 1 of the 95 IFA-negative samples was positive in the assay. These results strongly suggest that IgM capture ELISA using the recombinant Bor56 antigen is a reliable and detailed method for the detection of early O. tsutsugamushi infection.
机译：为了使斑疹伤寒与其他急性发热性疾病区分开来，快速可靠的血清学诊断很重要。我们开发了一种免疫球蛋白M（IgM）捕获酶联免疫吸附测定（ELISA）方法，用于诊断人类最近的em虫东方 O的生物素化重组56-kDa蛋白的使用。 tsutsugamushi Boryong，Bor56，在大肠杆菌中表达为具有麦芽糖结合蛋白的融合蛋白。在测试中，血清IgM抗体被包被在微量滴定板上的抗人IgM抗体捕获。通过向板中依次添加生物素化的Bor56抗原和过氧化物酶结合的链霉亲和素来揭示捕获的IgM抗体。通过检测确诊的立克次氏病患者和其他急性发热性疾病患者的176个血清样品，将IgM捕获ELISA与免疫荧光抗体测定（IFA）进行了比较。在这81个IgG IFA阳性样品中，有78个通过IgM捕获ELISA检测为阳性（灵敏度为96.3％），而所有31个IgM IFA阳性样品也都检测为阳性（灵敏度100％）。 IgM捕获ELISA的特异性为99％，并且95个IFA阴性样品中有1个在测定中为阳性。这些结果强烈表明，使用重组Bor56抗原的IgM捕获ELISA是检测早期 O的可靠且详细的方法。 tsutsugamushi 感染。

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