scholarly journals Epitope Mapping of the Immunodominant Invariable Region of Borrelia burgdorferi VlsE in Three Host Species

2000 ◽  
Vol 68 (4) ◽  
pp. 2349-2352 ◽  
Author(s):  
Fang Ting Liang ◽  
Mario T. Philipp
Keyword(s):  
Amino Acid ◽  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Immunosorbent Assay ◽  
Host Species ◽  
Antigenic Determinant ◽  
Epitope Mapping ◽  
Primate Species ◽  
Enzyme Linked Immunosorbent Assay ◽  
Variable Surface

ABSTRACT VlsE, the variable surface antigen of Borrelia burgdorferi, contains a 26-amino-acid-long immunodominant invariable region, IR6. In the present study, three overlapping 14-mer peptides reproducing the sequence of IR6were used as peptide-based enzyme-linked immunosorbent assay antigens to map this invariable region in infected monkeys, mice, and human Lyme disease patients. Antibodies of the two primate species appeared to recognize IR6 as a single antigenic determinant, while mouse antibodies recognized multiple epitopes within this region.

scholarly journals Sensitive and Specific Serodiagnosis of Lyme Disease by Enzyme-Linked Immunosorbent Assay with a Peptide Based on an Immunodominant Conserved Region of Borrelia burgdorferi VlsE

1999 ◽  
Vol 37 (12) ◽  
pp. 3990-3996 ◽  
Author(s):  
Fang Ting Liang ◽  
Allen C. Steere ◽  
Adriana R. Marques ◽  
Barbara J. B. Johnson ◽  
James N. Miller ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Immunosorbent Assay ◽  
Synthetic Peptide ◽  
Late Phase ◽  
Hospitalized Patients ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Variable Surface ◽  
Conserved Region

VlsE, the variable surface antigen of Borrelia burgdorferi, contains an immunodominant conserved region named IR6. In the present study, the diagnostic performance of a peptide enzyme-linked immunosorbent assay (ELISA) based on a 26-mer synthetic peptide (C6) with the IR6 sequence was explored. Sensitivity was assessed with serum samples (n = 210) collected from patients with clinically defined Lyme disease at the acute (early localized or early disseminated disease), convalescent, or late disease phase. The sensitivities for acute-, convalescent-, and late-phase specimens were 74% (29 of 39), 85 to 90% (34 of 40 to 35 of 39), and 100% (59 of 59), respectively. Serum specimens from early neuroborreliosis patients were 95% positive (19 of 20), and those from an additional group of patients with posttreatment Lyme disease syndrome yielded a sensitivity of 62% (8 of 13). To assess the specificity of the peptide ELISA, 77 serum samples from patients with other spirochetal or chronic infections, autoimmune diseases, or neurologic diseases and 99 serum specimens from hospitalized patients in an area where Lyme disease is not endemic were examined. Only two potential false positives from the hospitalized patients were found, and the overall specificity was 99% (174 of 176). Precision, which was assessed with a panel of positive and negative serum specimens arranged in blinded duplicates, was 100%. Four serum samples with very high anti-OspA antibody titers obtained from four monkeys given the OspA vaccine did not react with the C6 peptide. This simple, sensitive, specific, and precise ELISA may contribute to alleviate some of the remaining problems in Lyme disease serodiagnosis. Because of its synthetic peptide base, it will be inexpensive to manufacture. It also will be applicable to serum specimens from OspA-vaccinated subjects.

scholarly journals Characterization of a Borrelia burgdorferi VlsE Invariable Region Useful in Canine Lyme Disease Serodiagnosis by Enzyme-Linked Immunosorbent Assay

2000 ◽  
Vol 38 (11) ◽  
pp. 4160-4166 ◽  
Author(s):  
Fang Ting Liang ◽  
Richard H. Jacobson ◽  
Reinhard K. Straubinger ◽  
Amy Grooters ◽  
Mario T. Philipp
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Antibody Response ◽  
Immunosorbent Assay ◽  
Protein A ◽  
Immunoblot Analysis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Entire Study ◽  
Variable Surface

Sera collected from dogs experimentally infected withBorrelia burgdorferi by tick inoculation were analyzed for an antibody response to each of the six invariable regions (IRs; i.e., IR1 to IR6) of VlsE, the variable surface antigen of B. burgdorferi. Six synthetic peptides (C1 to C6), which reproduced the six IR sequences were used as peptide-based, enzyme-linked immunosorbent assay (ELISA) antigens. Two IRs, IR2 and IR6, were found to be immunodominant. Studies with serially collected serum samples from experimentally infected dogs revealed that the antibody response to IR6 appears earlier and is stronger than that to IR2. Thus, the IR6 sequence alone appeared to be sufficient for serodiagnosis. When C6 alone was used as antigen, the peptide-based ELISA was positive in 7 of 23 dogs (30%) as early as 3 weeks postinfection. All dogs (n = 33) became strongly positive 1 or 2 weeks later, and this response persisted for the entire study, which lasted for 69 weeks. Of 55 sera submitted by veterinarians from dogs suspected of having Lyme disease, 19 were also positive by the C6 ELISA, compared to 20 positives detected by immunoblot analysis using cultured B. burgdorferi lysates as antigen. The sensitivity of using C2 and C6 together for detecting specific antibody in both experimentally infected and clinically diagnosed dogs was not better than sensitivity with C6 alone, confirming that C6 suffices as a diagnostic probe. Moreover, the C6 ELISA yielded 100% specificity with serum samples collected from 70 healthy dogs, 14 dogs with infections other than B. burgdorferi, and 15 animals vaccinated with either outer surface protein A, whole-spirochete vaccines, or the common puppy-vaccines. Therefore, this C6 ELISA was both sensitive and specific for the serodiagnosis of canine Lyme disease and could be used with vaccinated dogs.

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

2007 ◽  
Vol 14 (7) ◽  
pp. 875-879 ◽  
Author(s):  
Maria J. C. Gomes-Solecki ◽  
Luciana Meirelles ◽  
John Glass ◽  
Raymond J. Dattwyler
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Immunosorbent Assay ◽  
Erythema Migrans ◽  
The United States ◽  
Enzyme Linked Immunosorbent Assay ◽  
Diagnostic Assay ◽  
Assay Sensitivity ◽  
Selection Of ◽  
Antigenic Epitopes

ABSTRACTIn the absence of erythema migrans, the basis for diagnosis of Lyme disease is the demonstration of an antibody response againstBorrelia burgdorferiin 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 infectingBorreliagenospecies. 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 ofBorreliaspp. is not optimal for use as a universal diagnostic assay for Lyme disease.

scholarly journals Effects of OspA Vaccination on Lyme Disease Serologic Testing

1999 ◽  
Vol 37 (11) ◽  
pp. 3718-3721 ◽  
Author(s):  
Maria E. Aguero-Rosenfeld ◽  
Janet Roberge ◽  
Carol A. Carbonaro ◽  
John Nowakowski ◽  
Robert B. Nadelman ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Disease Control ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serologic Testing ◽  
Control And Prevention ◽  
Centers For Disease Control

This study presents the effects of OspA vaccination on two-step testing for Borrelia burgdorferi antibodies. Although vaccinees developed enzyme-linked immunosorbent assay reactivity, immunoblots did not fulfill Centers for Disease Control and Prevention criteria for positivity. Furthermore, OspA reactivity did not interfere with interpretation of immunoblots with sera from patients who developed early Lyme disease despite vaccination.

scholarly journals Significantly Improved Accuracy of Diagnosis of Early Lyme Disease by Peptide Enzyme-Linked Immunosorbent Assay Based on the Borreliacidal Antibody Epitope of Borrelia burgdorferi OspC

2008 ◽  
Vol 15 (6) ◽  
pp. 981-985 ◽  
Author(s):  
Dean A. Jobe ◽  
Steven D. Lovrich ◽  
Krista E. Asp ◽  
Michelle A. Mathiason ◽  
Stephanie E. Albrecht ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Western Blot ◽  
Immunosorbent Assay ◽  
Antibody Test ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
C Terminus ◽  
Igm Elisa ◽  
Improved Accuracy

ABSTRACT Highly specific borreliacidal antibodies are induced by infection with Borrelia burgdorferi, and the immunodominant response during early Lyme disease is specific for an epitope within the 7 amino acids nearest the C terminus of OspC. We evaluated the ability of an enzyme-linked immunosorbent assay (ELISA) based on a synthetic peptide (OspC7) that matched the region to detect the response and compared the sensitivity during early Lyme disease to that for an FDA-approved Western blot. When the optical density value was adjusted to 98% specificity based on the results from testing normal or uncharacterized sera (n = 236) or sera from patients with blood factors or illnesses that commonly produce antibodies that cross-react with B. burgdorferi antigens (n = 77), 115 (73%) of 157 sera from patients likely to have early Lyme disease were positive for immunoglobulin M (IgM) antibodies and 17 (11%) also had IgG antibodies. In addition, the IgM ELISA reactivities and the titers of antibodies detected by a flow cytometric borreliacidal antibody test correlated closely (r = 0.646). Moreover, the IgM ELISA was significantly more sensitive (P < 0.001) than the Western blot procedure. The findings therefore confirmed that the peptide IgM ELISA detected OspC borreliacidal antibodies and provided strong evidence that the test can eliminate the necessity for confirming early Lyme disease by a supplementary test such as Western blotting.

scholarly journals Recombinant Chimeric Borrelia Proteins for Diagnosis of Lyme Disease

2000 ◽  
Vol 38 (7) ◽  
pp. 2530-2535 ◽  
Author(s):  
Maria J. C. Gomes-Solecki ◽  
John J. Dunn ◽  
Benjamin J. Luft ◽  
Jonathan Castillo ◽  
Daniel E. Dykhuizen ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chimeric Proteins ◽  
Whole Cell ◽  
Assay Optimization

Current serologic Lyme disease tests use whole borrelia cells as the source of antigen. These assays are difficult to standardize and to optimize for sensitivity and specificity. To help solve these problems, we constructed a library of recombinant chimeric proteins composed of portions of key antigens of Borrelia burgdorferi. These proteins were then used to develop an enzyme-linked immunosorbent assay. We compared our assay with the most sensitive of three whole-cell borrelia assays. We found that the recombinant assay could detect antibodies significantly better from early Lyme disease sera (P < 0.05), and had the same sensitivity for late Lyme disease sera, as the most sensitive whole-cell borrelia assay. On potentially cross-reactive sera, the recombinant assay was more specific, but not significantly so, than the best whole-cell borrelia assay. Optimization of the recombinant assay offers the potential for a significant improvement in both sensitivity and specificity.

scholarly journals Evaluation of the Recombinant VlsE-Based Liaison Chemiluminescence Immunoassay for Detection of Borrelia burgdorferi and Diagnosis of Lyme Disease

2008 ◽  
Vol 15 (12) ◽  
pp. 1796-1804 ◽  
Author(s):  
Thomas B. Ledue ◽  
Marilyn F. Collins ◽  
John Young ◽  
Martin E. Schriefer
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Viral Infections ◽  
Relative Sensitivity ◽  
The United States ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Chemiluminescence Immunoassay ◽  
Variable Surface ◽  
Relative Specificity

ABSTRACT Recent efforts to improve the serologic diagnosis of Lyme disease have included the use of a synthetic peptide (C6) that reproduces the sequence of invariable region 6 of VlsE, the variable surface antigen of Borrelia burgdorferi. In the present study, the diagnostic performance of DiaSorin's recombinant VlsE-based chemiluminescence immunoassay in 1,947 human serum samples was evaluated. Sensitivity was determined using two serum panels from the CDC. For panel I, we observed sensitivities of 68.4% and 75.6% for subjects with early, localized (n = 19) or disseminated (n = 41) disease, respectively. For panel II, we observed sensitivities of 61.5% and 100% for subjects with early (n = 26) or late-stage (n = 11) disease, respectively. We observed a specificity of 99.5% for healthy donors (n = 600) living either in regions of the United States where the disease is endemic or in regions where it is not endemic. Overall, specificity among 207 potentially cross-reactive sera from subjects who had other spirochetal infections, nonspirochetal infections including bacterial and viral infections, or autoimmune or neurologic disease; who were positive for rheumatoid factor or anti-mouse antibodies; or who had been previously vaccinated for Lyme disease was 93.7%. In a direct comparison of 1,038 prospectively collected samples for Lyme disease testing we observed a relative sensitivity of 70%, a relative specificity of 99.1%, and an overall agreement of 97.1% between the DiaSorin recombinant VlsE chemiluminescence immunoassay and the Immunetics peptide-based C6 enzyme-linked immunosorbent assay.

scholarly journals C-Terminal Invariable Domain of VlsE Is Immunodominant but Its Antigenicity Is Scarcely Conserved among Strains of Lyme Disease Spirochetes

2001 ◽  
Vol 69 (5) ◽  
pp. 3224-3231 ◽  
Author(s):  
Fang Ting Liang ◽  
Lisa C. Bowers ◽  
Mario T. Philipp
Keyword(s):  
Lyme Disease ◽  
Antibody Response ◽  
Human Serum ◽  
Synthetic Peptide ◽  
Species Complex ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Variable Surface ◽  
Entire Sequence ◽  
Carboxyl Terminal

ABSTRACT VlsE, the variable surface antigen of Borrelia burgdorferi, contains two invariable domains located at the amino and carboxyl terminal ends, respectively, and a central variable domain. In this study, both immunogenicity and antigenic conservation of the C-terminal invariable domain were assessed. Mouse antiserum to a 51-mer synthetic peptide (Ct) which reproduced the entire sequence of the C-terminal invariable domain of VlsE from B. burgdorferi strain B31 was reacted on immunoblots with whole-cell lysates extracted from spirochetes of 12 strains from the B. burgdorferi sensu lato species complex. The antiserum recognized only VlsE from strain B31, indicating that epitopes of this domain differed among these strains. When Ct was used as enzyme-linked immunosorbent assay (ELISA) antigen, all of the seven monkeys and six mice that were infected with B31 spirochetes produced a strong antibody response to this peptide, indicating that the C-terminal invariable domain is immunodominant. None of 12 monkeys and only 11 of 26 mice that were infected with strains other than B31 produced a detectable anti-Ct response, indicating a limited antigenic conservation of this domain among these strains. Twenty-six of 33 dogs that were experimentally infected by tick inoculation were positive by the Ct ELISA, while only 5 of 18 serum samples from dogs clinically diagnosed with Lyme disease contained detectable anti-Ct antibody. Fifty-seven of 64 serum specimens that were collected from American patients with Lyme disease were positive by the Ct ELISA, while only 12 of 21 European samples contained detectable anti-Ct antibody. In contrast, antibody to the more conserved invariable region IR6 of VlsE was present in all of these dog and human serum samples.

Epitope Mapping of an Anti-CD20 Monoclonal Antibody (C20Mab-60) Using Enzyme-Linked Immunosorbent Assay

2021 ◽  
Vol 40 (6) ◽  
pp. 250-254
Author(s):  
Junko Takei ◽  
Hiroyuki Suzuki ◽  
Teizo Asano ◽  
Guanjie Li ◽  
Masaki Saito ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Epitope Mapping ◽  
Enzyme Linked Immunosorbent Assay ◽  
Anti Cd20
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