Validity of Interpretation Criteria for Standardized Western Blots (Immunoblots) for Serodiagnosis of Lyme Borreliosis Based on Sera Collected throughout Europe

Abstract

Western blotting (WB; immunoblotting) is a widely used tool for the serodiagnosis of Lyme borreliosis (LB), but so far, no generally accepted criteria for performance and interpretation have been established in Europe. The current study was preceeded by a detailed analysis of WB with whole-cell lysates of three species ofBorrelia burgdorferi sensu lato (U. Hauser, G. Lehnert, R. Lobentanzer, and B. Wilske, J. Clin. Microbiol. 35:1433–1444, 1997). In that study, interpretation criteria for a positive WB result were developed with the data for 330 serum samples (from patients with LB in different stages [n = 189] and from a control group [n = 141]) originating mostly from southern Germany. In the present work, the interpretation criteria for strains PKo (Borrelia afzelii) and PBi (Borrelia garinii) developed in the previous study were reevaluated with 224 serum samples (from patients with LB in different stages [n = 97] and from a control group [n = 127]) originating from throughout Europe that were provided by the European Union Concerted Action on Lyme Borreliosis (EUCALB). De novo criteria were developed on the basis of the reactivities of the EUCALB sera and were evaluated with the data for the samples from southern Germany. Comparison of all results led to the following recommendations: For WB for immunoglobulin G (IgG), at least two bands among p83/100, p58, p43, p39, p30, OspC, p21, p17, and p14 for PKo and at least one band among p83/100, p39, p30, OspC, p21, and p17b for PBi; for WB for IgM, at least one band among p39, OspC, and p17 or a strong p41 band for PKo and at least one band among p39 and OspC or a strong p41 band for PBi. WB with PKo was the most sensitive, and this strain is recommended for use in WB for the serodiagnosis of LB throughout Europe.

Evidence of Dbps (decorin binding proteins) among European strains ofBorrelia burgdorferisensu lato and in the immune response of LB patient sera

“…Our data are the first report regarding immune responses in European patient sera to both Dbp proteins and give indirect evidence of their expression in Borrelia during human infection. The level of reactivity to Dbps that we found, mostly in early LB sera, was not as high as the level obtained with the mouse immune response, which is unanimously claimed to be strong and early against DbpA [3,4] and was lower compared to the high reactivity of p41, p58, p53, p21 OspC and p83–100 [5–8] reported in both early and late LB European human sera. Nevertheless we would like to point out that the reactivity we observed in immunoblots of patient sera in the 20–22 kDa region attributed to OspC and p21, is probably also due to the reactivity of Dbps.…”

To test or not to test? Laboratory support for the diagnosis of Lyme borreliosis: a position paper of ESGBOR, the ESCMID study group for Lyme borreliosis

“…p100, p17/p18) fits well with late manifestations (e.g. arthritis, acrodermatitis chronica atrophicans) [51]. Both patterns of antibody responses are also consistent with asymptomatic persistence of antibodies (past infection).…”

Comparison of isolation rate of Borrelia burgdorferi sensu lato in two different culture media, MKP and BSK-H

“…Isolation of B. burgdorferi sensu lato from clinical material represents the reference standard for demonstration of Borrelia infection and enables further molecular analysis of the isolated strains with a potential for enhancement of our knowledge on the relationships between Borrelia species, biological material (ticks, animals, humans) and clinical manifestations , as well as for the assessment of phenotypic and genotypic changes of the microorganism in different conditions . However, with the exception of skin specimens obtained from patients with cutaneous manifestations, the yield of this approach in European Lyme borreliosis is usually < 10% .…”