Evaluation of in vivo expressed Borrelia burgdorferi antigens for improved IgM serodiagnosis of early Lyme disease

Brandt, Kevin S.; Ullmann, Amy J.; Molins, Claudia R.; Horiuchi, Kalanthe; Biggerstaff, Brad J.; Gilmore, Robert D.

doi:10.1016/j.diagmicrobio.2018.09.012

Cited by 9 publications

(22 citation statements)

References 24 publications

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“…Additionally, IgM-based detection of some targets has demonstrated high sensitivity for early LD. These include targets used in the IgM Western blot (OspC, BmpA, P41) [14] and other novel targets (BBA65, BBA69, BBA70, BBA73) [42]. In a patient with early LD and negative STTT result, the GC-FP biochip detected IgG against BBA65 and BBA69, which agrees with a previous study that detected IgM antibodies against these two proteins [42].…”

Section: Gc-fp Analysis May Have Better Diagnostic Sensitivity Than Tsupporting

confidence: 89%

“…These include targets used in the IgM Western blot (OspC, BmpA, P41) [14] and other novel targets (BBA65, BBA69, BBA70, BBA73) [42]. In a patient with early LD and negative STTT result, the GC-FP biochip detected IgG against BBA65 and BBA69, which agrees with a previous study that detected IgM antibodies against these two proteins [42]. The addition of anti-IgM reporter antibody onto the same biochip increased the signal of these targets and led to detection of 8 other targets.…”

Section: Gc-fp Analysis May Have Better Diagnostic Sensitivity Than Tmentioning

confidence: 99%

“…We hypothesized that GC-FP analysis could be used to effectively: 1) screen for antibody targets relevant to various stages of LD, and 2) allow us to develop a new algorithm for diagnosing disease status that is more sensitive, specific, and informative than the STTT. Our efforts involved physical [39] ErpG, ErpY, ErpL OspE/F-like proteins [40] DbpA, DbpB Decorin-binding proteins [41] BBA65, BBA69, BBA70, BBA73 IgM reactivity in early Lyme disease [42] https://doi.org/10.1371/journal.pone.0228772.t001 assay development and characterization (i.e. determining the analytical sensitivity, resilience to changes in assay conditions, and variation across replicates), as well as generating a method to analyze the quantitative data (i.e.…”

Section: Introductionmentioning

confidence: 99%

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A fluorescent plasmonic biochip assay for multiplex screening of diagnostic serum antibody targets in human Lyme disease

et al. 2020

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Lyme disease (LD) diagnosis using the current two-tier algorithm is constrained by low sensitivity for early-stage infection and ambiguity in determining treatment response. We recently developed a protein microarray biochip that measures diagnostic serum antibody targets using grating-coupled fluorescent plasmonics (GC-FP) technology. This strategy requires microliters of blood serum to enable multiplexed biomarker screening on a compact surface and generates quantitative results that can be further processed for diagnostic scoring. The GC-FP biochip was used to detect serum antibodies in patients with active and convalescent LD, as well as various negative controls. We hypothesized that the quantitative, high-sensitivity attributes of the GC-FP approach permit: 1) screening of antibody targets predictive for LD status, and 2) development a diagnostic algorithm that is more sensitive, specific, and informative than the standard ELISA and Western blot assays. Notably, our findings led to a diagnostic algorithm that may be more sensitive than the current standard for detecting early LD, while maintaining 100% specificity. We further show that analysis of relative antibody levels to predict disease status, such as in acute and convalescent stages of infection, is possible with a highly sensitive and quantitative platform like GC-FP. The results from this study add to the urgent conversation regarding better diagnostic strategies and more effective treatment for patients affected by tick-borne disease.

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Section: Gc-fp Analysis May Have Better Diagnostic Sensitivity Than Tsupporting

confidence: 89%

Section: Gc-fp Analysis May Have Better Diagnostic Sensitivity Than Tmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A fluorescent plasmonic biochip assay for multiplex screening of diagnostic serum antibody targets in human Lyme disease

et al. 2020

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“…coli BL21 (DE3) according to the manufacturer’s instructions. Recombinant proteins were expressed and purified using a QiaExpress Ni-NTA FastStart kit (Qiagen, Valencia, CA) as described previously 39 . Proteins (100 ng/individual protein) were transferred to PVDF membranes for immunoblotting with mouse or human serum samples (1:200), followed by incubation with alkaline phosphatase conjugated goat anti-mouse or goat anti-human IgM or IgG (1:5000) with development by NBT/BCIP with similar approximated reaction times across all blots according to standard procedures.…”

Section: Methodsmentioning

confidence: 99%

Immunoproteomic analysis of Borrelia miyamotoi for the identification of serodiagnostic antigens

Harris

Harton

Marques

et al. 2019

Sci Rep

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The tick-borne spirochete, Borrelia miyamotoi, is an emerging pathogen of public health significance. Current B. miyamotoi serodiagnostic testing depends on reactivity against GlpQ which is not highly sensitive on acute phase serum samples. Additionally, anti-B. miyamotoi antibodies can cross-react with C6 antigen testing for B. burgdorferi, the causative agent of Lyme disease, underscoring the need for improved serological assays that produce accurate diagnostic results. We performed an immunoproteomics analysis of B. miyamotoi proteins to identify novel serodiagnostic antigens. Sera from mice infected with B. miyamotoi by subcutaneous inoculation or tick bite were collected for immunoblotting against B. miyamotoi membrane-associated proteins separated by 2-dimensional electrophoresis (2DE). In total, 88 proteins in 40 2DE immunoreactive spots were identified via mass spectrometry. Multiple variable large proteins (Vlps) and a putative lipoprotein were among those identified and analyzed. Reactivity of anti-B. miyamotoi sera against recombinant Vlps and the putative lipoprotein confirmed their immunogenicity. Mouse anti-B. burgdorferi serum was cross-reactive to all recombinant Vlps, but not against the putative lipoprotein by IgG. Furthermore, antibodies against the recombinant putative lipoprotein were present in serum from a B. miyamotoi-infected human patient, but not a Lyme disease patient. Results presented here provide a comprehensive profile of B. miyamotoi antigens that induce the host immune response and identify a putative lipoprotein as a potentially specific antigen for B. miyamotoi serodetection.

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“…The antigens BBA65, BBA70, and BBA73 were selected for IgM serum immunoreactivity evaluation in early Lyme disease patients together with the three antigens currently used in IgM second-tier immunoblotting, OspC, BmpA, and FlaB. We found that a six antigen approach, whereby reactivity against at least 2 of 6 antigens constituted a positive serology, could increase sensitivity without compromising specificity (6). Also in our initial screening of antigens, BBA69 and BBA73 demonstrated IgG reactivity in a set of early Lyme disease patient samples (5).…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Patient IgM and IgG Reactivity Against Multiple Antigens for Improvement of Serodiagnostic Testing for Early Lyme Disease

Brandt

Horiuchi

Biggerstaff

et al. 2019

Front. Public Health

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Serologic testing is the standard for laboratory diagnosis and confirmation of Lyme disease. Serodiagnostic assays to detect antibodies against Borrelia burgdorferi, the agent of Lyme borreliosis, are used for detection of infection. However, serologic testing within the first month of infection is less sensitive as patients' antibody responses continue to develop. Previously, we screened several B. burgdorferi in vivo expressed antigens for candidates that elicit early antibody responses in patients with Stage 1 and 2 Lyme disease. We evaluated patient IgM seroreactivity against 6 antigens and found an increase in sensitivity without compromising specificity when compared to current IgM second-tier immunoblot scoring. In this study, we continued the evaluation using a multi-antigen panel to measure IgM plus IgG seroreactivity in these early Lyme disease patients' serum samples. Using two statistical methods for calculating positivity cutoff values, sensitivity was 70 and 84–87%, for early acute and early convalescent Lyme disease patients, respectively. Specificity was 98–100% for healthy non-endemic control patients, and 96–100% for healthy endemic controls depending on the statistical analysis. We conclude that improved serologic testing for early Lyme disease may be achieved by the addition of multiple borrelial antigens that elicit IgM and IgG antibodies early in infection.

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Evaluation of in vivo expressed Borrelia burgdorferi antigens for improved IgM serodiagnosis of early Lyme disease

Cited by 9 publications

References 24 publications

A fluorescent plasmonic biochip assay for multiplex screening of diagnostic serum antibody targets in human Lyme disease

A fluorescent plasmonic biochip assay for multiplex screening of diagnostic serum antibody targets in human Lyme disease

Immunoproteomic analysis of Borrelia miyamotoi for the identification of serodiagnostic antigens

Evaluation of Patient IgM and IgG Reactivity Against Multiple Antigens for Improvement of Serodiagnostic Testing for Early Lyme Disease

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