Wilske Bettina
Max von Pettenkofer Institute, University of Munich, National Reference Center for Borreliae, Pettenkofer-Stresse 9a, D 80336 Munich, Germany.
Vector Borne Zoonotic Dis. 2003 Winter;3(4):215-27. doi: 10.1089/153036603322662200.
In Europe, Lyme borreliosis is caused by at least three species, B. burgdorferi sensu stricto, B. afzelii and B. garinii. Thus microbiological diagnosis in European patients must consider the heterogeneity of Lyme disease borreliae for development of diagnostic tools such as PCR primers and diagnostic antigens. According to guidelines of the German Society of Hygiene and Microbiology, the serological diagnosis should follow the principle of a two-step procedure. A sensitive ELISA differentiating IgM and IgG is recommended as the first step. In case the ELISA is reactive, it is followed by immunoblots (IgM and IgG) as the second step. The reactive diagnostic bands should be clearly identified, which is easy if recombinant antigens are used. The sensitivity and standardization of immunoblots has been considerably enhanced by use of recombinant antigens instead of whole cell lysates. Improved sensitivity resulted from use of recombinant proteins that are expressed primarily in vivo (e.g., VlsE) and combination of homologous proteins from different strains of borrelia (e.g., DbpA). It also appears promising to use recombinant proteins (DbpA, VlsE, others) or synthetic peptides (the conserved C6 peptide derived from VlsE) as ELISA antigens. At present, detection rates for serum antibodies are 20-50% in stage I, 70-90% in stage II, and nearly 100% in stage III Lyme disease. The main goals for the future are to improve specificity in general and sensitivity for diagnosis of early manifestations (stage I and II). Detection of the etiological agent by culture or PCR should be confined to specific indications and specialised laboratories. Recommended specimens are skin biopsy specimens, CSF and synovial fluid. The best results are obtained from skin biopsies with culture or PCR (50-70%) and synovial tissue or fluid (50-70% with PCR). CSF yields positive results in only 10-30% of patients. Methods that are not recommended for diagnostic purposes are antigen tests in body fluids, PCR of urine, and lymphocyte transformation tests.
在欧洲，莱姆病疏螺旋体病至少由三种菌株引起，即狭义伯氏疏螺旋体、阿氏疏螺旋体和伽氏疏螺旋体。因此，对于欧洲患者的微生物学诊断，在开发诸如聚合酶链反应（PCR）引物和诊断抗原等诊断工具时，必须考虑莱姆病疏螺旋体的异质性。根据德国卫生与微生物学会的指南，血清学诊断应遵循两步法原则。第一步推荐使用能区分IgM和IgG的敏感酶联免疫吸附测定（ELISA）。如果ELISA呈阳性反应，则第二步进行免疫印迹法（IgM和IgG）检测。应明确识别出呈阳性反应的诊断条带，如果使用重组抗原则很容易做到这一点。与使用全细胞裂解物相比，使用重组抗原极大地提高了免疫印迹法的灵敏度和标准化程度。使用主要在体内表达的重组蛋白（如VlsE）以及来自不同疏螺旋体菌株的同源蛋白组合（如DbpA）提高了灵敏度。将重组蛋白（DbpA、VlsE等）或合成肽（源自VlsE的保守C6肽）用作ELISA抗原似乎也很有前景。目前，莱姆病I期血清抗体的检出率为20% - 50%，II期为70% - 90%，III期接近100%。未来的主要目标是总体上提高特异性以及早期表现（I期和II期）诊断的灵敏度。通过培养或PCR检测病原体应限于特定指征和专业实验室。推荐的标本是皮肤活检标本、脑脊液和滑液。皮肤活检进行培养或PCR可获得最佳结果（50% - 70%），滑膜组织或滑液进行PCR也可获得较好结果（50% - 70%）。脑脊液仅在10% - 30%的患者中呈阳性结果。不推荐用于诊断目的的方法包括体液中的抗原检测、尿液PCR以及淋巴细胞转化试验。
