Development of a Metabolic Biosignature for Detection of Early Lyme Disease

Molins, Claudia R.; Ashton, Laura V.; Wormser, Gary P.; Hess, Ann; Delorey, Mark J.; Mahapatra, Sebabrata; Schriefer, Martin E.; Belisle, John T.

doi:10.1093/cid/civ185

Cited by 70 publications

(99 citation statements)

References 37 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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“…From the perspective of the use of disease-related biomarkers, approaches involving metabolomic and immunologic profiles have shown promise (22)(23)(24). In particular, a liquid chromatography-mass spectrometry-based analysis of metabolites was shown to correctly classify early-phase Lyme disease patients and healthy controls with a sensitivity of 88%.…”

Section: Discussionmentioning

confidence: 99%

“…In particular, a liquid chromatography-mass spectrometry-based analysis of metabolites was shown to correctly classify early-phase Lyme disease patients and healthy controls with a sensitivity of 88%. Given that the test is not specific to B. burgdorferi, a remarkable specificity of 95% was achieved in tests against "look-alike diseases" such as fibromyalgia and rheumatoid arthritis (22). However, patients with late-stage disease were not tested and the potential for the levels of biomarkers included in the profile to wane over time is a concern.…”

Section: Discussionmentioning

confidence: 99%

Five-Antigen Fluorescent Bead-Based Assay for Diagnosis of Lyme Disease

Embers

Hasenkampf

Barnes

et al. 2016

Clin Vaccine Immunol

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The systematically difficult task of diagnosing Lyme disease can be simplified by sensitive and specific laboratory tests. The currently recommended two-tier test for serology is highly specific but falls short in sensitivity, especially in the early acute phase. We previously examined serially collected serum samples from Borrelia burgdorferi-infected rhesus macaques and defined a combination of antigens that could be utilized for detection of infection at all phases of disease in humans. The five B. burgdorferi antigens, consisting of OspC, OspA, DbpA, OppA2, and the C6 peptide, were combined into a fluorescent cytometric beadbased assay for the detection of B. burgdorferi antigen-specific IgG antibodies. Samples from Lyme disease patients and controls were used to determine the diagnostic value of this assay. Using this sample set, we found that our five-antigen multiplex IgG assay exhibited higher sensitivity (79.5%) than the enzyme immunoassay (EIA) (76.1%), the two-tier test (61.4%), and the C6 peptide enzyme-linked immunosorbent assay (ELISA) (77.2%) while maintaining specificity over 90%. When detection of IgM was added to the bead-based assay, the sensitivity improved to 91%, but at a cost of reduced specificity (78%). These results indicate that the rational combination of antigens in our multiplex assay may offer an improved serodiagnostic test for Lyme disease. With an estimated 300,000 new cases each year, Lyme disease is the most common vector-borne disease in North America (http://www.cdc.gov/lyme/stats/humanCases.html) (1). The burden of this disease in many parts of Europe is also staggering (1, 2). Treatment with antibiotics is generally effective, even more so when employed soon after infection (3). Therefore, early and reliable laboratory diagnosis is critical for effective cure of Lyme disease patients.Patient serum antibodies specific for Borrelia burgdorferi antigens are detected with currently recommended laboratory tests for Lyme disease. While the detection of antigen may be preferred for early diagnosis, this is encumbered by the absence of detectable spirochetes or spirochetal antigen in the bloodstream once the organism has disseminated. Thus, the use of antigen detection from a blood or skin biopsy specimen has not demonstrated favorable sensitivity (4). The two most commonly used tests for diagnosis of Lyme disease in North America are (i) the two-tier test that includes an enzyme-linked immunosorbent assay (ELISA) and confirmatory Western blotting and (ii) the C6 test, where antibodies to a specific peptide within a conserved region of VlsE, the B. burgdorferi antigen, are detected (5-8). Both the twotier and C6 tests exhibit high specificity and are most sensitive for patients in the disseminated phases of disease (5, 8-10). In addition, the C6 test has been evaluated as an indicator of treatment outcome in the United States (11, 12), with a Ն4-fold decline in antibody titer (up to 6 months after treatment) in a majority of patients. The Western blot procedure is technically dema...

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“…Using liquid chromatography mass spectrometry and statistical modeling, Molins et al defined a metabolic profile of 44 biosignatures, primarily lipid and lipophilic molecules, present in patients with early LD and compared the performance of this panel with that of the traditional and 2-ELISA TTTAs. The metabolomics assay outperformed both TTTAs with respect to sensitivity (88% versus 43% to 48%, respectively; P Ͻ 0.0001) and showed statistically equivalent specificities among healthy controls and individuals with a non-LD condition (29). Although certain challenges with the methodology exist, on the basis of the preliminary data, metabolomic profiling appears to hold promise as a future tool for the diagnosis of LD.…”

Section: Novel Approaches For Ld Detectionmentioning

confidence: 91%

“…Finally, as an alternative to classic antibody detection for LD, the CDC, along with multiple academic and clinical collaborators, recently evaluated the applicability of metabolomics, defined as the evaluation of low-molecular-weight (Ͻ1,500-Da) biomolecules, for the diagnosis of early LD (29). An individual's baseline metabolic profile can be altered by a variety of environmental stressors, and the detection of changes to this baseline may lay the foundation for the development of a novel diagnostic tool for infections, including LD.…”

Section: Novel Approaches For Ld Detectionmentioning

confidence: 99%

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

Theel

2016

J Clin Microbiol

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Lyme disease prevails as the most commonly transmitted tick-borne infection in the United States, and serologic evaluation for antibodies to Borrelia burgdorferi remains 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.

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Development of a Metabolic Biosignature for Detection of Early Lyme Disease

Abstract: The data provide proof-of-concept that metabolic profiling for early Lyme disease can achieve significantly greater (P < .0001) diagnostic sensitivity than current 2-tier serology, while retaining high specificity.

Cited by 70 publications

References 37 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

Five-Antigen Fluorescent Bead-Based Assay for Diagnosis of Lyme Disease

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

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