Can ELISPOT Be Applied to A Clinical Setting as A Diagnostic Utility for Neuroborreliosis?

Nordberg, Marika; Forsberg, Pia; Nyman, Dag; Skogman, Barbro H.; Nyberg, Clara; Ernerudh, Jan; Eliasson, Ingvar; Ekerfelt, Christina

doi:10.3390/cells1020153

Cited by 22 publications

(14 citation statements)

References 48 publications

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“…High specificity was obtained using a two-tier test with the first-stage test optical density threshold lowered. Nordberg et al63 was excluded as this study had a specificity of 82%; the sensitivity was also low at 36% and would have depressed the study mean sensitivity. Craven et al’s52 study had a low specificity of 71% with a sensitivity of 51%, which would not significantly bias the overall results of this study.…”

Section: Resultsmentioning

confidence: 99%

Commercial test kits for detection of Lyme borreliosis: a meta-analysis of test accuracy

Cook¹,

Puri²

2016

IJGM

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The clinical diagnosis of Lyme borreliosis can be supported by various test methodologies; test kits are available from many manufacturers. Literature searches were carried out to identify studies that reported characteristics of the test kits. Of 50 searched studies, 18 were included where the tests were commercially available and samples were proven to be positive using serology testing, evidence of an erythema migrans rash, and/or culture. Additional requirements were a test specificity of ≥85% and publication in the last 20 years. The weighted mean sensitivity for all tests and for all samples was 59.5%. Individual study means varied from 30.6% to 86.2%. Sensitivity for each test technology varied from 62.4% for Western blot kits, and 62.3% for enzyme-linked immunosorbent assay tests, to 53.9% for synthetic C6 peptide ELISA tests and 53.7% when the two-tier methodology was used. Test sensitivity increased as dissemination of the pathogen affected different organs; however, the absence of data on the time from infection to serological testing and the lack of standard definitions for “early” and “late” disease prevented analysis of test sensitivity versus time of infection. The lack of standardization of the definitions of disease stage and the possibility of retrospective selection bias prevented clear evaluation of test sensitivity by “stage”. The sensitivity for samples classified as acute disease was 35.4%, with a corresponding sensitivity of 64.5% for samples from patients defined as convalescent. Regression analysis demonstrated an improvement of 4% in test sensitivity over the 20-year study period. The studies did not provide data to indicate the sensitivity of tests used in a clinical setting since the effect of recent use of antibiotics or steroids or other factors affecting antibody response was not factored in. The tests were developed for only specific Borrelia species; sensitivities for other species could not be calculated.

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Section: Resultsmentioning

confidence: 99%

Commercial test kits for detection of Lyme borreliosis: a meta-analysis of test accuracy

Cook¹,

Puri²

2016

IJGM

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“…Borrelia -specific T cell immunity has not yet been studied sufficiently due to the lack of highly sensitive and specific T cell-based assays that would be suited for the clinical laboratory. Several attempts have been made to study T cell reactivity against Borrelia , but the results were not consistent from different studies [18,19,20]. There is increasing evidence, however, that T cell assays have potential advantages over antibody-based assays in the detection of Borrelia infections.…”

Section: Introductionmentioning

confidence: 99%

An Enhanced ELISPOT Assay for Sensitive Detection of Antigen-Specific T Cell Responses to Borrelia burgdorferi

Jin¹,

Roen²,

Lehmann

et al. 2013

Cells

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Lyme Borreliosis is an infectious disease caused by the spirochete Borrelia burgdorferi that is transmitted through the bite of infected ticks. Both B cell-mediated humoral immunity and T cell immunity develop during natural Borrelia infection. However, compared with humoral immunity, the T cell response to Borrelia infection has not been well elucidated. In this study, a novel T cell-based assay was developed and validated for the sensitive detection of antigen-specific T cell response to B. burgdorferi. Using interferon-γ as a biomarker, we developed a new enzyme-linked immunospot method (iSpot LymeTM) to detect Borrelia antigen-specific effector/memory T cells that were activated in vivo by exposing them to recombinant Borrelia antigens ex vivo. To test this new method as a potential laboratory diagnostic tool, we performed a clinical study with a cohort of Borrelia positive patients and healthy controls. We demonstrated that the iSpot Lyme assay has a significantly higher specificity and sensitivity compared with the Western Blot assay that is currently used as a diagnostic measure. A comprehensive evaluation of the T cell response to Borrelia infection should, therefore, provide new insights into the pathogenesis, diagnosis, treatment and monitoring of Lyme disease.

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“…Von Baehr et al [13] evaluated three Borrelia species and did not find any difference. Nordberg et al [44] used B. garinii as a stimulating agent, CSF instead of blood and nitrocellulose-bottomed ELISpot plates instead of PVDF-bottomed plates, and also did not find higher numbers of activated T cells in their ELISpot assay. The Osp-mix we used contained antigens derived from an LNB-associated strain (B. garinii); however, the Osp-mix was inferior compared to the use of B. burgdorferi B31, as discussed in the previous paragraph.…”

Section: Discussionmentioning

confidence: 99%

Prospective comparison of two enzyme-linked immunosorbent spot assays for the diagnosis of Lyme neuroborreliosis

Gorkom

Voet

Sankatsing

et al. 2020

Clinical and Experimental Immunology

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Summary Commercial cellular tests are used to diagnose Lyme borreliosis (LB), but studies on their clinical validation are lacking. This study evaluated the utility of an in‐house and a commercial enzyme‐linked immunosorbent spot (ELISpot) assay for the diagnosis of Lyme neuroborreliosis (LNB). Prospectively, peripheral blood mononuclear cells (PBMCs) were isolated from patients and controls and analysed using an in‐house Borrelia ELISpot assay and the commercial LymeSpot assay. B. burgdorferi B31 whole cell lysate and a mixture of outer surface proteins were used to stimulate the PBMCs and the numbers of interferon‐gamma‐secreting T cells were measured. Results were evaluated using receiver operating characteristic (ROC) curve analysis. Eighteen active and 12 treated LNB patients, 10 healthy individuals treated for an early (mostly cutaneous) manifestation of LB in the past and 47 untreated healthy individuals were included. Both assays showed a poor diagnostic performance with sensitivities, specificities, positive and negative predictive values ranging from 44.4–66.7%, 42.0–72.5%, 21.8–33.3% and 80.5–87.0%, respectively. The LymeSpot assay performed equally poorly when the calculation method of the manufacturer was used. Both the in‐house and the LymeSpot assay are unable to diagnose active LNB or to monitor antibiotic treatment success.

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Can ELISPOT Be Applied to A Clinical Setting as A Diagnostic Utility for Neuroborreliosis?

Cited by 22 publications

References 48 publications

Commercial test kits for detection of Lyme borreliosis: a meta-analysis of test accuracy

Commercial test kits for detection of Lyme borreliosis: a meta-analysis of test accuracy

An Enhanced ELISPOT Assay for Sensitive Detection of Antigen-Specific T Cell Responses to Borrelia burgdorferi

Prospective comparison of two enzyme-linked immunosorbent spot assays for the diagnosis of Lyme neuroborreliosis

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