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莱姆病血清学检测的局限性:酶联免疫吸附测定(ELISA)和免疫印迹法与聚合酶链反应(PCR)及培养方法的比较评估

Limitation of serological testing for Lyme borreliosis: evaluation of ELISA and western blot in comparison with PCR and culture methods.

作者信息

Tylewska-Wierzbanowska Stanislawa, Chmielewski Tomasz

机构信息

Department of Bacteriology, National Institute of Hygiene, Warsaw, Poland.

出版信息

Wien Klin Wochenschr. 2002 Jul 31;114(13-14):601-5.

Abstract

The aim of the study was to evaluate a one-step procedure using an ELISA test of high specificity and a two-step procedure using immunoblot as a confirmation test, and to compare the results of serological testing with detection of bacterial DNA and living spirochetes. Sera, synovial (SF) and cerebro-spinal fluids (CSF) were obtained from 90 patients with clinical symptoms of Lyme borreliosis. Serum samples were tested with recombinant ELISA and Western blot assay. Citrated blood, cerebrospinal and synovial fluids samples were cultured in cell line and tested by PCR to detect spirochetes. No correlation was found between levels of specific B. burgdorferi antibodies detected with a recombinant antigen ELISA and the number of protein fractions developed with these antibodies by immunoblot. Moreover, Lyme borreliosis patients who have live spirochetes in body fluids have low or negative levels of borrelial antibodies in their sera. This indicates that an efficient diagnosis of Lyme borreliosis has to be based on a combination of various techniques such as serology, PCR and culture, not solely on serology.

摘要

本研究的目的是评估使用高特异性酶联免疫吸附测定(ELISA)试验的一步法以及使用免疫印迹作为确认试验的两步法,并将血清学检测结果与细菌DNA和活螺旋体的检测结果进行比较。从90例有莱姆病临床症状的患者中获取血清、滑膜液(SF)和脑脊液(CSF)。血清样本用重组ELISA和蛋白质印迹法检测。枸橼酸盐抗凝血、脑脊液和滑膜液样本在细胞系中培养,并用聚合酶链反应(PCR)检测螺旋体。用重组抗原ELISA检测的特异性伯氏疏螺旋体抗体水平与这些抗体通过免疫印迹产生的蛋白条带数量之间未发现相关性。此外,体液中有活螺旋体的莱姆病患者血清中伯氏疏螺旋体抗体水平较低或为阴性。这表明莱姆病的有效诊断必须基于血清学、PCR和培养等多种技术的联合应用,而不能仅依赖血清学。

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