Comparative evaluation of three different ELISA methods for the diagnosis of early culture-confirmed Lyme disease in Italy

Lyme borreliosis (LB) is a disease for which antibody-based detection assays are often required for diagnosis. The variable surface molecule VlsE and IR6, one of its invariable regions, are commonly targeted by the antibody response in infected individuals. A series of enzyme-linked immunosorbent assays was performed to comparatively examine the antibody responses of North American LB patients (n ‫؍‬ 37) to VlsE and invariable segments of this molecule. Both immunoglobulin M (IgM) and IgG responses to full-length VlsE and to peptides reproducing invariable regions 2, 4, and 6, as well as the invariable domains at the amino and carboxyl termini of VlsE, were assessed. The proportions and specificities of reactivity to the invariable segments were tested by using cognate peptides as competitors for VlsE binding by patient serum antibodies. IR6 epitopes (by the C6 peptide) were found to dominate the response to invariable segments. IR6 (C6)-specific antibodies were detected in 78% of the serum specimens, whereas <40% of patients generated antibodies that bound the N-or C-terminal domain and <12% of patients responded to either IR2 or IR4. Interestingly, 15 of 37 patients generated IgG antibodies that reacted with C6 but not with VlsE. Conversely, IgM responses were frequent for VlsE but not for invariable segments. A representative number of the serum specimens (n ‫؍‬ 8) that contained IgG antibodies reacting with both C6 and VlsE was assessed in competition experiments, using C6 as a competitor. Only half of these specimens contained IgG antibodies whose binding to VlsE could be inhibited >50% by competition with the added C6 peptide. The median percent inhibition was 45.5%. These findings indicate that IR6 epitopes are largely concealed from the VlsE molecular surface and that full-length VlsEbased diagnosis likely detects antibodies to conformational and/or variable region epitopes.

Section: Discussionmentioning

confidence: 99%

Dominant Epitopes of the C6 Diagnostic Peptide ofBorrelia burgdorferiAre Largely Inaccessible to Antibody on the Parent VlsE Molecule

Embers

Jacobs

Johnson

et al. 2007

“…The C6 assay has become the assay of choice both in the United States and Europe, either alone (3,6,8,9,14,17) or in combination with other assays (1,4,12). Sensitivity of this assay in Europe varies between 91% in a study in The Netherlands against a panel of sera from clinically defined Lyme patients confirmed by immunoblotting (16) and 33% in Italy against a panel of sera from clinically defined Lyme patients with EM confirmed by culture (10).…”

Section: Discussionmentioning

confidence: 99%

Epitope Length, Genospecies Dependency, and Serum Panel Effect in the IR6 Enzyme-Linked Immunosorbent Assay for Detection of Antibodies toBorrelia burgdorferi

Gomes-Solecki

Meirelles

Glass

et al. 2007

In the absence of erythema migrans, the basis for diagnosis of Lyme disease is the demonstration of an antibody response against Borrelia burgdorferi in an appropriate clinical setting. The C6 enzyme-linked immunosorbent assay, based on the IR6 region of VlsE, has become widely used in both the United States and Europe. We mapped the antigenic epitopes of IR6 to a shorter sequence that is equivalent in sensitivity and specificity to the full-length IR6 25-residue peptide. In addition, we observed significant differences in sensitivity between serum panels (60 to 100%), indicating that the selection of the serum panels can shape the apparent overall sensitivity of the assay. Contrary to prior reports, the assay sensitivity is greater when the IR6 peptide is derived from the sequence of the same infecting Borrelia genospecies. Using our North American panels and the two panels obtained from European Lyme disease patients, we determined that the IR6 assay that is based on a single genospecies of Borrelia spp. is not optimal for use as a universal diagnostic assay for Lyme disease.

“…In Europe, a few papers on the diagnostic use of the C6 peptide have been published (9,14,16,22,23). Moreover, Bacon and coworkers (2) reported that the test with the C6 peptide performed as well as or better than the two-tiered algorithm.…”

mentioning

confidence: 99%

A Decrease in the Immunoglobulin G Antibody Response against the VlsE Protein of Borrelia burgdorferi Sensu Lato Correlates with the Resolution of Clinical Signs in Antibiotic-Treated Patients with Early Lyme Disease

Marangoni

Sambri

Accardo

et al. 2006

Self Cite

The purpose of this study was to evaluate the diagnostic performance of the LIAISON Borrelia Screen (Diasorin, Saluggia, Italy), a new automated immunoassay based on the chemiluminescent technology (chemiluminescence immunoassay). To assess whether a decrease in a negative value in the anti-VlsE immunoglobulin G (IgG) antibody titer was correlated with a positive response to treatment, a group of serially collected serum samples from 67 patients with culture-confirmed erythema migrans was retrospectively studied. All the patients had been treated with antibiotics and were free of disease within 3 to 6 months of follow-up. All the 15 patients who were found to be IgG positive at the time of enrollment and who were bled at least four times during the follow-up became IgG seronegative at 2 to 6 months posttreatment. These results indicate that a decline in the anti-VlsE antibody titer coincides with effective antimicrobial therapy in patients with early localized Lyme disease.