Decorin Binding Proteins A and B in the Serodiagnosis of Lyme Disease in North America

Arnaboldi, Paul M.; Sambir, Mariya; Dattwyler, Raymond J.

doi:10.1128/cvi.00383-14

Cited by 18 publications

(24 citation statements)

References 30 publications

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“…Although novel approaches for the diagnosis of Lyme disease are currently being developed that include serology-based and non-serology-based assays (12)(13)(14)(15), for now, serologic testing, and specifically an STTT algorithm using an EIA (or IFA), followed by immunoblotting, remains the CDC-recommended diagnostic testing method for patients with Lyme disease (2). The findings presented here demonstrate the strengths and weaknesses of STTT using well-characterized serum samples and controls and also support previous findings that a 2-EIA-based MTTT algo-rithm for serologic diagnostic testing for Lyme disease may be a beneficial addition to the currently recommended laboratory diagnostic algorithm for Lyme disease.…”

Section: Discussionmentioning

confidence: 99%

Lyme Borreliosis Serology: Performance of Several Commonly Used Laboratory Diagnostic Tests and a Large Resource Panel of Well-Characterized Patient Samples

et al. 2016

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The current recommendation for the laboratory confirmation of Lyme disease is serology-based diagnostics. Specifically, a standardized two-tiered testing (STTT) algorithm is applied that utilizes a first-tier immunofluorescence assay or enzyme immunoassay (EIA) that, if the result is positive or equivocal, is followed by second-tier immunoblotting. Despite the standardization and performance achievements, STTT is considered technically complex and subjective, as well as insensitive for early acute infection. These issues have prompted development of novel algorithms and testing platforms. In this study, we evaluated the performance of several commonly used assays for STTT. Several modified two-tiered testing (MTTT) algorithms, including a 2-EIA algorithm and modified criteria for second-tier IgG immunoblots, were also evaluated. All tests were performed on sera from a recently available, well-defined archive of positive-and negative-control patients. Our study demonstrates differences in the results between individual first-and second-tier tests, although the overall agreement of the different STTT algorithms used was strong. In addition, the MTTT algorithm utilizing 2-EIAs was found to be equivalent to all STTT algorithms tested, with agreement ranging from 94 to 97%. The 2-EIA MTTT algorithm slightly enhanced sensitivity in early disease compared to the STTT algorithms evaluated. Furthermore, these data add to the mounting evidence that a 2-EIA-based MTTT algorithm, where immunoblotting is replaced by the C6 EIA, performs as well or better than STTT. Serologic testing has been the mainstay of laboratory diagnostics for Lyme disease for over 20 years. To date in the United States, serologic tests are the only U.S. Food and Drug Administration (FDA) 510(k)-cleared diagnostic tests for Lyme disease (1). Standardization of serologic testing for Lyme disease occurred in 1994, when attendees of the Second National Conference on Serologic Diagnosis of Lyme Disease (Dearborn, MI) determined that a two-tiered algorithm, optimized for specificity and the highest sensitivity, was most appropriate for Lyme disease laboratory diagnosis (2). The standard two-tiered testing (STTT) algorithm utilizes a first-tier immunofluorescence assay (IFA) or enzyme immunoassay (EIA) that, if the result is positive or equivocal, is followed by a second-tier immunoblot (IgM and/or IgG). The interpretation criteria for a positive immunoblot requires that at least two of three or at least five of ten bands be considered positive for IgM and IgG reactivity, respectively. Further, it was recommended that patients only be tested by IgM immunoblotting if their duration of illness is 30 days or less. STTT and interpretation criteria continue to be recommended by the Centers for Disease Control and Prevention (CDC) for laboratory confirmation of Lyme disease (2).Several dozen individual first-and second-tier diagnostic tests have been FDA 510(k) cleared and are commercially available. Most first-tier tests use either Borrelia burgdorferi whole-cel...

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Section: Discussionmentioning

confidence: 99%

Lyme Borreliosis Serology: Performance of Several Commonly Used Laboratory Diagnostic Tests and a Large Resource Panel of Well-Characterized Patient Samples

et al. 2016

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“…HNCACB, CBCACONH, HNCOCA, and HNCO experiments were collected for backbone atom assignment. For side chain atom assignments, CCONH, HCCONH, HCCH-TOCSY, 15 N-edited TOCSY, and 13 C-and 15 N-edited NOESY-HSQC spectra were collected. Both HN and NC RDCs were collected using J-modulated pulse sequence on both proteins aligned in 6% polyacrylamide gel.…”

Section: Acquisition and Analysis Of Nmr Structural Data For N40 And mentioning

confidence: 99%

“…Unlabeled protein was grown in normal LB while isotopically labeled protein was grown in M9 medium supplemented with 15 NH 4 Cl or [ 13 C] glucose. The bacteria were induced with 0.5 mM IPTG and incubated overnight at 30°C.…”

Section: Experimental Expression and Purification Of B31 N40 And Pbmentioning

confidence: 99%

“…In particular, because of DBPA's strong tendency to induce human immune response, it has been considered as a component in vaccines and serological tests for Lyme disease. [15] However, the large sequence variation in DBPA may require the inclusion of several versions of DBPA to elicit comprehensive immunity against all strains since sequence variations in DBPA may prevent antibodies against DBPA of one strain from detecting other versions. Understanding of the structural differences and similarities among the different strains will aid in the intelligent selection of representative DBPA variants that can present greatest coverage in these tests.…”

Section: Introductionmentioning

confidence: 99%

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Structural mechanisms underlying sequence-dependent variations in GAG affinities of decorin binding protein A, a Borrelia burgdorferi adhesin

Morgan

Wang

2015

Biochemical Journal

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Abbreviations: DBPA, decorin-binding protein A. DBPB, decorin-binding protein B. GAG, glycosaminoglycan. DS, dermatan sulfate. HSQC, heteronuclear single quantum coherence spectroscopy.Summary statement: Structural mechanisms behind variations in GAG affinities of DBPAs from different Borrelia strains were investigated using NMR. DBPA from strain PBr was revealed to have an additional GAG-binding epitope and a retracted linker allowing more access to its GAG-binding sites. AbstractDecorin binding protein A (DBPA) is an important surface adhesin of the bacterium Borrelia burgdorferi, the causative agent of Lyme disease. DBPA facilitates the bacteria's colonization of human tissue by adhering to glycosaminoglycan (GAG), a sulfated polysaccharide. Interestingly, DBPA sequence variation among different strains of Borrelia spirochetes is high, resulting in significant differences in their GAG affinities. However, the structural mechanisms contributing to these differences are unknown. We determined the solution structures of DBPAs from strain N40 of Borrelia burgdorferi and strain PBr of Borrelia gariini, two DBPA variants whose GAG affinities deviate significantly from strain B31, the best characterized version of DBPA. Our structures revealed that significant differences exist between PBr DBPA and B31/N40 DBPAs. In particular, the C-terminus of PBr DBPA, unlike C-termini from B31 and N40 DBPAs, is positioned away from the GAG-binding pocket, and the linker between helices one and two of PBr DBPA is highly structured and retracted from the GAGbinding pocket. The repositioning of the C-terminus allowed the formation of an extra GAGbinding epitope in PBr DBPA, and the retracted linker give GAG ligands more access to the GAG-binding epitopes than other DBPAs. Characterization of GAG ligands' interactions with wild type PBr and mutants confirmed the importance of the second major GAG-binding epitope and established the fact that the two epitopes are independent of one another and the new epitope is as important to GAG binding as the traditional epitope.

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“…Epitope mapping of B. burgdorferi surface proteins has revealed novel peptides conserved across B. burgdorferi genospecies that are not found in other bacterial species (18). Synthetic antigen peptides derived from these epitopes demonstrate equivalent sensitivity and decreased cross-reactivity (19,20).…”

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confidence: 99%

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

et al. 2015

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iThe current standard for laboratory diagnosis of Lyme disease in the United States is serologic detection of antibodies against Borrelia 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 62 B. burgdorferi surface 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 early B. burgdorferi infection 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.

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Decorin Binding Proteins A and B in the Serodiagnosis of Lyme Disease in North America

Cited by 18 publications

References 30 publications

Lyme Borreliosis Serology: Performance of Several Commonly Used Laboratory Diagnostic Tests and a Large Resource Panel of Well-Characterized Patient Samples

Lyme Borreliosis Serology: Performance of Several Commonly Used Laboratory Diagnostic Tests and a Large Resource Panel of Well-Characterized Patient Samples

Structural mechanisms underlying sequence-dependent variations in GAG affinities of decorin binding protein A, a Borrelia burgdorferi adhesin

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

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