Diagnostic Value of Proteins of ThreeBorreliaSpecies (Borrelia burgdorferiSensu Lato) and Implications for Development and Use of Recombinant Antigens for Serodiagnosis of Lyme Borreliosis in Europe

Abstract

More and more assays for the serodiagnosis of Lyme borreliosis (LB) are based on recombinant antigens. However, so far, there is no consensus as to which are the most specific and sensitive proteins and how they should be used in combination to obtain tests with the best discrimination abilities. The present study was preceded by a detailed analysis of Western blots (WB) using whole-cell lysates ofBorrelia burgdorferi sensu stricto strain PKa2, B. afzelii PKo, and B. garinii PBi (U. Hauser, G. Lehnert, R. Lobentanzer, and B. Wilske, J. Clin. Microbiol. 35:1433–1444, 1997). For the present work, the data bank from that study, containing information about the reactivities of 330 sera (from patients at different stages of LB [n = 189]; control group, n = 141), was reused. The specificities and sensitivities of various combinations of proteins from different strains were calculated for different interpretation criteria. For immunoglobulin G (IgG) WB, the recommended combination of antigens available to date as recombinant proteins included p83/100 of PKa2, p83/100 of PKo, p39 of PKo, p39 of PBi, and OspC of PBi (interpretation criterion, at least one reactive band required for a positive WB; specificity, 96.5%; sensitivity, 56.1%). The further addition of p58 of PKo, p17 of PKo, or p14 of PKo was most favorable in terms of both a considerable gain of sensitivity and little loss of specificity. IgG Western blotting with a whole-cell lysate of strain PKo might be improved by the addition of OspC of PBi. For IgG WB, the best combination, out of all bands, was p83/100, p58, p39, p30, and p21 of all three strains and OspC of PBi, p17b of PBi, p56 of PKa2, p43 of PKo, p17 of PKo, and p14 of PKo (interpretation criterion, at least two reactive bands required for a positive WB; specificity, 97.2%; sensitivity, 61.4%). An interpretation criterion of at least two reactive bands is more reliable than one of only one reactive band. For IgM WB, the best combination was OspC of PKo, OspC of PBi, p39 of all three strains, p17 of PKo, and strong reactions with p41 of all three strains (interpretation criterion, at least one reactive band required; specificity, 97.9%; sensitivity, 47.0%).

Interannual Dynamics of Antibody Levels in Naturally Infected Long-Lived Colonial Birds

“…The most prevalent immunoblot bands corresponded to highly specific antigens of Borrelia burgdorferi s.l. : p58, B. garinii OspC and p39 (Hauser et al 1998). The comparison of the immunoblots with Ab levels showed that individuals with high OD values for the ELISA (.0.62) had IgY Ab against at least three of the diagnostic Borrelia antigens, the criterion for positive diagnosis in humans (Wilske 2003).…”

“…We used immunoblots (Western blots) to confirm that high OD values meant that the individuals had been exposed to antigens of Borrelia burgdorferi sensu lato (Wilske 2003). The immunoblots also enabled us to characterize the immunological profile of the Borrelia antigens against which Ab were produced (Hauser et al 1998, Pachner et al 2002 and, consequently, to investigate the repeatability of banding patterns between years (10 seropositive individuals sampled in 2005 and 2006). The immunoblot assays were performed using a commercial kit (Western Blot Lyme IgG þ VlsE, Meridian Bioscience Europe, Nice, France).…”

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

“…Methods. All methods have previously been described in detail (15,16), and only brief descriptions will be given here.…”

Microbiological and serological diagnosis of Lyme borreliosis

“…strain PKo) (Wilske et al, 2000). The immunoblot criteria recommended by the Centers for Disease Control (CDC) that are used in the USA are not applicable for European patients (Hauser et al, 1997(Hauser et al, , 1998Robertson et al, 2000). Dressler et al (1994) have shown, in an immunoblot study, that the immune response of European patients is restricted to a narrower spectrum of Borrelia proteins, compared with that shown by American patients.…”