scholarly journals Development of a Multiantigen Panel for Improved Detection of Borrelia burgdorferi Infection in Early Lyme Disease

2015 ◽  
Vol 53 (12) ◽  
pp. 3834-3841 ◽  
Author(s):  
Lauren J. Lahey ◽  
Michael W. Panas ◽  
Rong Mao ◽  
Michelle Delanoy ◽  
John J. Flanagan ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Early Stage ◽  
Erythema Migrans ◽  
Disease Patient ◽  
High Specificity ◽  
The United States ◽  
Surface Proteins ◽  
Current Standard ◽  
Content Type

The current standard for laboratory diagnosis of Lyme disease in the United States is serologic detection of antibodies againstBorrelia burgdorferi. The Centers for Disease Control and Prevention recommends a two-tiered testing algorithm; however, this scheme has limited sensitivity for detecting early Lyme disease. Thus, there is a need to improve diagnostics for Lyme disease at the early stage, when antibiotic treatment is highly efficacious. We examined novel and established antigen markers to develop a multiplex panel that identifies early infection using the combined sensitivity of multiple markers while simultaneously maintaining high specificity by requiring positive results for two markers to designate a positive test. Ten markers were selected from our initial analysis of 62B. burgdorferisurface proteins and synthetic peptides by assessing binding of IgG and IgM to each in a training set of Lyme disease patient samples and controls. In a validation set, this 10-antigen panel identified a higher proportion of early-Lyme-disease patients as positive at the baseline or posttreatment visit than two-tiered testing (87.5% and 67.5%, respectively;P< 0.05). Equivalent specificities of 100% were observed in 26 healthy controls. Upon further analysis, positivity on the novel 10-antigen panel was associated with longer illness duration and multiple erythema migrans. The improved sensitivity and comparable specificity of our 10-antigen panel compared to two-tiered testing in detecting earlyB. burgdorferiinfection indicates that multiplex analysis, featuring the next generation of markers, could advance diagnostic technology to better aid clinicians in diagnosing and treating early Lyme disease.

scholarly journals Antibodies to Borrelia burgdorferi OspA, OspC, OspF, and C6 Antigens as Markers for Early and Late Infection in Dogs

2012 ◽  
Vol 19 (4) ◽  
pp. 527-535 ◽  
Author(s):  
Bettina Wagner ◽  
Heather Freer ◽  
Alicia Rollins ◽  
David Garcia-Tapia ◽  
Hollis N. Erb ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
The United States ◽  
Sensu Stricto ◽  
Antibody Responses ◽  
Surface Proteins ◽  
Early Infection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Content Type ◽  
Antibody Levels

ABSTRACTLyme disease in the United States is caused byBorrelia burgdorferisensu stricto, which is transmitted to mammals by infected ticks.Borreliaspirochetes differentially express immunogenic outer surface proteins (Osp). Our aim was to evaluate antibody responses to Osp antigens to aid the diagnosis of early infection and the management of Lyme disease. We analyzed antibody responses during the first 3 months after the experimental infection of dogs using a novel multiplex assay. Results were compared to those obtained with two commercial assays detecting C6 antigen. Multiplex analysis identified antibodies to OspC and C6 as early as 3 weeks postinfection (p.i.) and those to OspF by 5 weeks p.i. Antibodies to C6 and OspF increased throughout the study, while antibodies to OspC peaked between 7 and 11 weeks p.i. and declined thereafter. A short-term antibody response to OspA was observed in 3/8 experimentally infected dogs on day 21 p.i. Quant C6 enzyme-linked immunosorbent assay (ELISA) results matched multiplex results during the first 7 weeks p.i.; however, antibody levels subsequently declined by up to 29%. Immune responses then were analyzed in sera from 125 client-owned dogs and revealed high agreement between antibodies to OspF and C6 as robust markers for infection. Results from canine patient sera supported that OspC is an early infection marker and antibodies to OspC decline over time. The onset and decline of antibody responses toB. burgdorferiOsp antigens and C6 reflect their differential expression during infection. They provide valuable tools to determine the stage of infection, treatment outcomes, and vaccination status in dogs.

scholarly journals Outer Surface Protein C Peptide Derived from Borrelia burgdorferi Sensu Stricto as a Target for Serodiagnosis of Early Lyme Disease

2013 ◽  
Vol 20 (4) ◽  
pp. 474-481 ◽  
Author(s):  
Paul M. Arnaboldi ◽  
Rudra Seedarnee ◽  
Mariya Sambir ◽  
Steven M. Callister ◽  
Josephine A. Imparato ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Bacterial Species ◽  
Erythema Migrans ◽  
Early Infection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Attractive Alternative ◽  
Serum Samples ◽  
Peptide Epitope ◽  
Content Type

ABSTRACTCurrent serodiagnostic assays for Lyme disease are inadequate at detecting early infection due to poor sensitivity and nonspecificity that arise from the use of whole bacteria or bacterial proteins as assay targets; both targets contain epitopes that are cross-reactive with epitopes found in antigens of other bacterial species. Tests utilizing peptides that contain individual epitopes highly specific forBorrelia burgdorferias diagnostic targets are an attractive alternative to current assays. Using an overlapping peptide library, we mapped linear epitopes in OspC, a critical virulence factor ofB. burgdorferirequired for mammalian infection, and confirmed the results by enzyme-linked immunosorbent assay (ELISA). We identified a highly conserved 20-amino-acid peptide epitope, OspC1. Via ELISA, OspC1 detected specific IgM and/or IgG in 60 of 98 serum samples (62.1%) obtained from patients with erythema migrans (early Lyme disease) at the time of their initial presentation. By comparison, the commercially available OspC peptide PepC10 detected antibody in only 48 of 98 serum samples (49.0%). In addition, OspC1 generated fewer false-positive results among negative healthy and diseased (rheumatoid arthritis and positive Rapid Plasma Reagin [RPR+] test result) control populations than did PepC10. Both highly specific and more sensitive than currently available OspC peptides, OspC1 could have value as a component of a multipeptide Lyme disease serological assay with significantly improved capabilities for the diagnosis of early infection.

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 The Past, Present, and (Possible) Future of Serologic Testing for Lyme Disease

2016 ◽  
Vol 54 (5) ◽  
pp. 1191-1196 ◽  
Author(s):  
Elitza S. Theel
Keyword(s):  
United States ◽  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Metabolic Profiling ◽  
Historical Perspective ◽  
The United States ◽  
Performance Characteristics ◽  
Content Type ◽  
The Past ◽  
Testing Algorithm

Lyme disease prevails as the most commonly transmitted tick-borne infection in the United States, and serologic evaluation for antibodies toBorrelia burgdorferiremains the recommended modality for diagnosis. This review presents a brief historical perspective on the evolution of serologic assays for Lyme disease and provides a summary of the performance characteristics for the currently recommended two-tiered testing algorithm (TTTA). Additionally, a recently proposed alternative to the traditional TTTA is discussed, and novel methodologies, including immuno-PCR and metabolic profiling for Lyme disease, are outlined.

scholarly journals Completed Genome Sequences of Borrelia burgdorferiSensu Stricto B31(NRZ) and Closely Related Patient Isolates from Europe

2017 ◽  
Vol 5 (28) ◽  
Author(s):  
Gabriele Margos ◽  
Sabrina Hepner ◽  
Christoph Mang ◽  
Andreas Sing ◽  
Bernhard Liebl ◽  
...  
Keyword(s):  
United States ◽  
Borrelia Burgdorferi ◽  
Lyme Borreliosis ◽  
Causative Agent ◽  
Erythema Migrans ◽  
The United States ◽  
Sensu Stricto ◽  
Genome Sequences ◽  
Content Type

ABSTRACT Borrelia burgdorferi sensu stricto is a causative agent of human Lyme borreliosis in the United States and Europe. We report here the completed genome sequences of strain B31 isolated from a tick in the United States and two closely related strains from Europe, PAli and PAbe, which were isolated from patients with erythema migrans and neuroborreliosis, respectively.

scholarly journals Single Core Genome Sequencing for Detection of both Borrelia burgdorferi Sensu Lato and Relapsing Fever Borrelia Species

2019 ◽  
Vol 16 (10) ◽  
pp. 1779
Author(s):  
Sin Hang Lee ◽  
John Eoin Healy ◽  
John S Lambert
Keyword(s):  
United States ◽  
Lyme Disease ◽  
16S Rrna ◽  
Borrelia Burgdorferi ◽  
Genome Sequencing ◽  
Core Genome ◽  
Direct Detection ◽  
Early Stage ◽  
The United States ◽  
Relapsing Fever

Lyme disease, initially described as Lyme arthritis, was reported before nucleic-acid based detection technologies were available. The most widely used diagnostic tests for Lyme disease are based on the serologic detection of antibodies produced against antigens derived from a single strain of Borrelia burgdorferi. The poor diagnostic accuracy of serological tests early in the infection process has been noted most recently in the 2018 Report to Congress issued by the U.S. Department of Health and Human Services Tick-Borne Disease Working Group. Clinical Lyme disease may be caused by a diversity of borreliae, including those classified as relapsing fever species, in the United States and in Europe. It is widely accepted that antibiotic treatment of Lyme disease is most successful during this critical early stage of infection. While genomic sequencing is recognized as an irrefutable direct detection method for laboratory diagnosis of Lyme borreliosis, development of a molecular diagnostic tool for all clinical forms of borreliosis is challenging because a “core genome” shared by all pathogenic borreliae has not yet been identified. After a diligent search of the GenBank database, we identified two highly conserved segments of DNA sequence among the borrelial 16S rRNA genes. We further developed a pair of Borrelia genus-specific PCR primers for amplification of a segment of borrelial 16S rRNA gene as a “core genome” to be used as the template for routine Sanger sequencing-based metagenomic direct detection test. This study presented examples of base-calling DNA sequencing electropherograms routinely generated in a clinical diagnostic laboratory on DNA extracts of human blood specimens and ticks collected from human skin bites and from the environment. Since some of the tick samples tested were collected in Ireland, borrelial species or strains not known to exist in the United States were also detected by analysis of this 16S rRNA “core genome”. We recommend that hospital laboratories located in Lyme disease endemic areas begin to use a “core genome” sequencing test to routinely diagnose spirochetemia caused by various species of borreliae for timely management of patients at the early stage of infection.

scholarly journals Evaluation of Selected Borrelia burgdorferi lp54 Plasmid-Encoded Gene Products Expressed during Mammalian Infection as Antigens To Improve Serodiagnostic Testing for Early Lyme Disease

2015 ◽  
Vol 22 (11) ◽  
pp. 1176-1186 ◽  
Author(s):  
Zachary P. Weiner ◽  
Rebecca M. Crew ◽  
Kevin S. Brandt ◽  
Amy J. Ullmann ◽  
Martin E. Schriefer ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Recombinant Proteins ◽  
Erythema Migrans ◽  
Disease Patient ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Early Stages ◽  
Lyme Carditis ◽  
Mammalian Infection

ABSTRACTLaboratory testing for the diagnosis of Lyme disease is performed primarily by serologic assays and is accurate for detection beyond the acute stage of the infection. Serodiagnostic assays to detect the early stages of infection, however, are limited in their sensitivity, and improvement is warranted. We analyzed a series ofBorrelia burgdorferiproteins known to be induced within feeding ticks and/or during mammalian infection for their utility as serodiagnostic markers against a comprehensive panel of Lyme disease patient serum samples. The antigens were assayed for IgM and IgG reactivity in line immunoblots and separately by enzyme-linked immunosorbent assay (ELISA), with a focus on reactivity against early Lyme disease with erythema migrans (EM), early disseminated Lyme neuroborreliosis, and early Lyme carditis patient serum samples. By IgM immunoblotting, we found that recombinant proteins BBA65, BBA70, and BBA73 reacted with early Lyme EM samples at levels comparable to those of the OspC antigen used in the current IgM blotting criteria. Additionally, these proteins reacted with serum samples from patients with early neuroborreliosis and early carditis, suggesting value in detecting early stages of this disease progression. We also found serological reactivity against recombinant proteins BBA69 and BBA73 with early-Lyme-disease samples using IgG immunoblotting and ELISA. Significantly, some samples that had been scored negative by the Centers for Disease Control and Prevention-recommended 2-tiered testing algorithm demonstrated positive reactivity to one or more of the antigens by IgM/IgG immunoblot and ELISA. These results suggest that incorporating additionalin vivo-expressed antigens into the current IgM/IgG immunoblotting tier in a recombinant protein platform assay may improve the performance of early-Lyme-disease serologic testing.

scholarly journals Human Antibody Responses to VlsE Antigenic Variation Protein of Borrelia burgdorferi

1999 ◽  
Vol 37 (12) ◽  
pp. 3997-4004 ◽  
Author(s):  
M. B. Lawrenz ◽  
J. M. Hardham ◽  
R. T. Owens ◽  
J. Nowakowski ◽  
A. C. Steere ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Lupus Erythematosus ◽  
Antigenic Variation ◽  
Erythema Migrans ◽  
Disease Patient ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Antibody ◽  
Whole Cell ◽  
Cell Elisa

VlsE is a 35-kDa surface-exposed lipoprotein of Borrelia burgdorferi that was shown previously to undergo antigenic variation through segmental recombination of silent vlscassettes with vlsE during experimental mouse infections. Previous data had indicated that sera from North American Lyme disease patients and experimentally infected animals contained antibodies reactive with VlsE. In this study, sera from patients with Lyme disease, syphilis, and autoimmune conditions as well as from healthy controls were examined for reactivity with VlsE by Western blotting and enzyme-linked immunosorbent assay (ELISA). Strong Western blot reactivity to a recombinant VlsE cassette region protein was obtained consistently with Lyme disease sera. Although sera from Lyme disease patients also reacted with a band corresponding to VlsE in B. burgdorferi B31-5A3, interpretation was complicated by low levels of VlsE expression in in vitro-cultured B. burgdorferi and by the presence of comigrating bands. An ELISA using recombinant VlsE was compared with an ELISA using sonically disrupted B. burgdorferi as the antigen. For a total of 93 Lyme disease patient sera examined, the VlsE ELISA yielded sensitivities of 63% for culture-confirmed erythema migrans cases and 92% for later stages, as compared to 61 and 98%, respectively, for the “whole-cell” ELISA. The specificities of the two assays with healthy blood donor sera were comparable, but the VlsE ELISA was 90% specific with sera from syphilis patients, compared to 20% specificity for the whole-cell ELISA with this group. Neither assay showed reactivity with a panel of sera from 20 non-Lyme disease arthritis patients or 20 systemic lupus erythematosus patients. Our results indicate that VlsE may be useful in the immunodiagnosis of Lyme disease and may offer greater specificity than ELISAs using whole B. burgdorferi as the antigen.

scholarly journals The Lyme Disease Biobank: Characterization of 550 Patient and Control Samples from the East Coast and Upper Midwest of the United States

2020 ◽  
Vol 58 (6) ◽  
Author(s):  
Elizabeth J. Horn ◽  
George Dempsey ◽  
Anna M. Schotthoefer ◽  
U. Lena Prisco ◽  
Matthew McArdle ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Laboratory Testing ◽  
Erythema Migrans ◽  
Public Health Problem ◽  
The United States ◽  
Content Type ◽  
Institutional Review ◽  
Testing Algorithm ◽  
And Control ◽  
Positive Results

ABSTRACT Lyme disease (LD) is an increasing public health problem. Current laboratory testing is insensitive in early infection, the stage at which appropriate treatment is most effective in preventing disease sequelae. The Lyme Disease Biobank (LDB) collects samples from individuals with symptoms consistent with early LD presenting with or without erythema migrans (EM) or an annular, expanding skin lesion and uninfected individuals from areas of endemicity. Samples were collected from 550 participants (298 cases and 252 controls) according to institutional review board-approved protocols and shipped to a centralized biorepository. Testing was performed to confirm the presence of tick-borne pathogens by real-time PCR, and a subset of samples was tested for Borrelia burgdorferi by culture. Serology was performed on all samples using the CDC’s standard two-tiered testing algorithm (STTTA) for LD. LD diagnosis was supported by laboratory testing in 82 cases, including positive results by use of the STTTA, PCR, or culture or positive results by two enzyme-linked immunosorbent assays for cases presenting with EM lesion sizes of >5 cm. The remaining 216 cases had negative laboratory testing results. For the controls, 43 were positive by at least one of the tiers and 6 were positive by use of the STTTA. The results obtained with this collection highlight and reinforce the known limitations of serologic testing in early LD, with only 29% of individuals presenting with EM lesion sizes of >5 cm yielding a positive result using the STTTA. Aliquots of whole blood, serum, and urine from clinically characterized patients with and without LD are available to investigators in academia and industry for evaluation or development of novel diagnostic assays for LD, to continue to improve upon currently available methods.

scholarly journals Discovery of the Lyme Disease Agent

mBio ◽  
2019 ◽  
Vol 10 (5) ◽  
Author(s):  
Alan G. Barbour ◽  
Jorge L. Benach
Keyword(s):  
Lyme Disease ◽  
Scientific Discovery ◽  
Long Island ◽  
Erythema Migrans ◽  
Rocky Mountain ◽  
The United States ◽  
Content Type ◽  
History Of ◽  
Disease Agent ◽  
Lyme Disease Agent

ABSTRACT A detailed first-hand account of the events leading up to the discovery of the Lyme disease agent has been lacking. Nearly 40 years have elapsed since the discovery of the organism that was named Borrelia burgdorferi. There are thousands of articles in the scientific and medical literature on this organism and the disease that it causes. In the interval since the organism’s discovery, however, misconceptions have arisen regarding not only the disease but the discovery itself. Accordingly, with this paper, we aim to fill in the details of this episode in medical history with a joint introduction, first-person accounts by the two authors, a summary of contemporaneous events, and concluding comments. The history of the discovery of the Lyme disease agent has threads originating in different places in the United States. Studies on Long Island, NY, provided the epidemiological thread of studies on rickettsial diseases and babesiosis, linking the latter with the cutaneous manifestation of Lyme disease, now known as erythema migrans. The Long Island thread intersected Montana’s Rocky Mountain Laboratories thread of studies on a relapsing fever Borrelia and its cultivation and expertise in vector biology. This intersection made possible the discovery of the spirochete and its recovery from patients. This paper stresses that what may seem to have been an individual scientific discovery is actually the product of several threads coming together and is attributable to more people than appreciated.

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