Evaluation of a lyme disease enzyme immunoassay using the 41-G fragment of flagellin

Abstract

An immunogenic region of the Borrelia burgdorferi flagellin encompassing amino acids 197-273 and designated 41-G was evaluated as an antigen in an enzyme immunoassay (EIA) for Lyme disease on a routine basis in a reference laboratory. Sera that tested positive for Lyme disease by EIA using 41-G or the whole-cell Borrelia burgdorferi lysate as the antigen were also evaluated by immunoblot for reactivity with Borrelia burgdorferi, and the patient's clinical history was determined retrospectively by a questionnaire distributed to the referring physician. The sensitivity of the 41-G based EIA for the serologic diagnosis of Lyme disease, when compared with that of the Borrelia burgdorferi lysate EIA, was 70% (35 of 50). These data demonstrate that 41-G has utility as an antigen in EIA, although the sensitivity is at present less than that of the assay employing the Borrelia burgdorferi whole-cell lysate.

Development of enzyme-linked immunosorbent assays using recombinant borrelial antigens for serodiagnosis of Borrelia burgdorferi infection

“…This corresponds well with previous Western blot and ELISA results in Lyme borreliosis patients [11,35,45]. It is known from former investigations characterizing several immunogenic regions of flagellin that the de-gree of sensitivity and specificity differs among the various flagellin fragments [6,16,26].…”

“…This has been confirmed by several clinical studies [11,22,31,35,45], indicating that the use of a single recombinant antigen is not sufficient for serodiagnosis of Lyme borreliosis. Our investigations with p100 revealed only a slightly lower test sensitivity when the assay was compared with the OGP-ELISA.…”

Enzyme-linked immunosorbent assays with recombinant internal flagellin fragments derived from different species of Borrelia burgdorferi sensu lato for the serodiagnosis of Lyme neuroborreliosis

“…Due to the strong reactivity of several sera (IFA confirmed) we assumed that they might represent true positive results and, therefore, the cut off absorbance values were defined by the 92th percentile. If sera from patients with all stages and manifestations of LB must be assessed in a routine setting a test based on a single antigen might not be as sensitive as an assay using a wholecell preparation or a combination of recombinant antigens [8,32,40]. However, the situation might be somewhat different for the serodiagnosis of acute NB.…”

Presentation of Peptide Antigens as Albumin Conjugates for Use in Detection of Serum Antibodies by Enzyme-Linked Immunosorbent Assay

“…In our preliminary serological test for Lyme disease, using a single epitope from Bb flagellin as the antigen, a sensitivity of 69% and a specificity of 96 or 91% were obtained for panels A and B, respectively. It should be noted that a similar flagellin epitope sequence was presented as a glutathione transferase fusion protein, and a similar sensitivity for detection of Lyme disease was reported (23). A parallel comparison of this set of serum samples with a commercial ELISA kit (MarDx Diagnostics, Carlsbad, CA), using Bb whole cell lysate as the antigen, gave a higher sensitivity (81 and 88%) but a lower specificity (68 and 71%).…”

“…It should be noted that a similar flagellin epitope sequence was presented as a glutathione transferase fusion protein, and a similar sensitivity for detection of Lyme disease was reported (23). A parallel comparison of this set of serum samples with a commercial ELISA kit (MarDx Diagnostics, Carlsbad, CA), using Bb whole cell lysate as the antigen, gave a higher sensitivity (81 and 88%) but a lower specificity (68 and 71%).…”