Interpretation criteria for standardized Western blots for three European species of Borrelia burgdorferi sensu lato.

Journal of Clinical Microbiology
U HauserB Wilske

Abstract

Western blots (WBs; immunoblots) are a widely used tool for the serodiagnosis of Lyme borreliosis, but so far, no defined criteria for performance, analysis, and interpretation have been established in Europe. For the current study WBs were produced with strains PKa2 (Borrelia burgdorferi sensu stricto), PKo (Borrelia afzelii), and PBi (Borrelia garinii). To improve resolution we used gels of 17 cm in length. In a first step, 13 immunodominant proteins were identified with monoclonal antibodies. Then, the apparent molecular masses of all visually distinguishable bands were determined densitometrically. Approximately 40 bands of between 14 and 100 kDa were differentiated for each strain. From a study with 330 serum samples (from 189 patients with Lyme borreliosis and 141 controls), all observed bands were documented. To establish criteria for a positive WB result, the discriminating ability of a series of band combinations (interpretation rules) were evaluated separately for each strain (for immunoglobulin G [IgG] WB, > 40 combinations; for IgM WB, > 15 combinations). The following interpretation criteria resulting in specificities of greater than 96% were recommended: for IgG WB, at least one band of p83/100, p58, p56, OspC, p2...Continue Reading

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