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来自狭义伯氏疏螺旋体、阿氏疏螺旋体和伽氏疏螺旋体的重组OspC在莱姆病血清学诊断中的应用

Recombinant OspC from Borrelia burgdorferi sensu stricto, B. afzelii and B. garinii in the serodiagnosis of Lyme borreliosis.

作者信息

Panelius Jaana, Lahdenne Pekka, Heikkilä Tero, Peltomaa Miikka, Oksi Jarmo, Seppälä Ilkka

机构信息

*Haartman Institute, Department of Bacteriology and Immunology, †Hospital for Children and Adolescents, ‡Department of Otorhinolaryngology, University of Helsinki, §Department of Medicine, Turku University Central Hospital, Turku and ∥HUCH Laboratory Diagnostics, Helsinki University Central Hospital, Helsinki, Finland.

出版信息

J Med Microbiol. 2002 Sep;51(9):731-739. doi: 10.1099/0022-1317-51-9-731.

Abstract

Genes for the outer-surface protein C (OspC) from three north European human isolates of Borrelia burgdorferi sensu stricto, B. afzelii and B. garinii were cloned and sequenced. Polyhistidine-tagged recombinant OspC (rOspC) proteins were produced in Escherichia coli and used, after biotinylation, as antigens on streptavidin-coated plates in enzyme-linked immunosorbent assays (ELISA). In IgM ELISA, 30% (5/17) and 35% (6/17) of patients with erythema migrans (EM) in the acute or convalescent phase, respectively, reacted with one to three rOspCs. Of the patients, 53% (8/15) with neuroborreliosis (NB) and 53% (8/15) with Lyme arthritis (LA) had IgM antibodies to OspC. The immunoreactivity was stronger against rOspC from B. afzelii and B. garinii than against rOspC from B. burgdorferi sensu stricto. In early Lyme borreliosis (LB), rOspC and flagella performed equally well in detecting IgM antibodies. Cross-reactive antibodies to rOspC were observed in serum samples from patients with rheumatoid factor positivity and with syphilis or Epstein-Barr virus (EBV) infection. In IgM ELISA, thiocyanate in the serum dilution buffer reduced EBV-associated non-specific positive reactions. Of the patient sera examined in IgG ELISA, 30% (5/17) with EM in the acute phase, 35% (6/17) with EM in the convalescent phase, 33% (5/15) with NB and 60% (9/15) with LA were positive. Because of the heterogeneity of OspC, a polyvalent antigen with several OspC variants from at least B. afzelii and B. garinii is needed to improve the sensitivity of OspC ELISA in the serodiagnosis of LB in Europe.

摘要

对来自北欧的三株狭义伯氏疏螺旋体、阿氏疏螺旋体和伽氏疏螺旋体的人类分离株的外表面蛋白C(OspC)基因进行了克隆和测序。在大肠杆菌中产生了多组氨酸标签的重组OspC(rOspC)蛋白,并在生物素化后,用作酶联免疫吸附测定(ELISA)中链霉亲和素包被板上的抗原。在IgM ELISA中,急性期或恢复期的游走性红斑(EM)患者分别有30%(5/17)和35%(6/17)与一至三种rOspC发生反应。患有神经型莱姆病(NB)的患者中有53%(8/15)以及患有莱姆关节炎(LA)的患者中有53%(8/15)具有针对OspC的IgM抗体。与来自狭义伯氏疏螺旋体的rOspC相比,针对来自阿氏疏螺旋体和伽氏疏螺旋体的rOspC的免疫反应性更强。在早期莱姆病(LB)中,rOspC和鞭毛在检测IgM抗体方面表现同样出色。在类风湿因子阳性以及患有梅毒或爱泼斯坦-巴尔病毒(EBV)感染的患者血清样本中观察到了针对rOspC的交叉反应性抗体。在IgM ELISA中,血清稀释缓冲液中的硫氰酸盐减少了EBV相关的非特异性阳性反应。在IgG ELISA检测的患者血清中,急性期EM患者中有30%(5/17)、恢复期EM患者中有35%(6/17)、NB患者中有33%(5/15)以及LA患者中有60%(9/15)呈阳性。由于OspC的异质性,需要一种包含至少来自阿氏疏螺旋体和伽氏疏螺旋体的几种OspC变体的多价抗原,以提高OspC ELISA在欧洲莱姆病血清诊断中的敏感性。

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