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Large differences between test strategies for the detection of anti-Borrelia antibodies are revealed by comparing eight ELISAs and five immunoblots.

Abstract

We investigated the influence of assay choice on the results in a two-tier testing algorithm for the detection of anti-Borrelia antibodies. Eighty-nine serum samples from clinically well-defined patients were tested in eight different enzyme-linked immunosorbent assay (ELISA) systems based on whole-cell antigens, whole-cell antigens supplemented with VlsE and assays using exclusively recombinant proteins. A subset of samples was tested in five immunoblots: one whole-cell blot, one whole-cell blot supplemented with VlsE and three recombinant blots. The number of IgM- and/or IgG-positive ELISA results in the group of patients suspected of Borrelia infection ranged from 34 to 59%. The percentage of positives in cross-reactivity controls ranged from 0 to 38%. Comparison of immunoblots yielded large differences in inter-test agreement and showed, at best, a moderate agreement between tests. Remarkably, some immunoblots gave positive results in samples that had been tested negative by all eight ELISAs. The percentage of positive blots following a positive ELISA result depended heavily on the choice of ELISA-immunoblot combination. We conclude that the assays used to detect anti-Borrelia antibodies have widely divergent sensitivity and specificity. The choice of ELISA-immunoblot combination severely influences the number of positive results, making the exchange of test results between laboratories with different methodologies hazardous.

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  • Authors+Show Affiliations

    ,

    VUMC, Amsterdam, The Netherlands. w.ang@vumc.nl

    , , ,

    Source

    MeSH

    Antibodies, Bacterial
    Antigens, Bacterial
    Borrelia
    Clinical Laboratory Techniques
    Enzyme-Linked Immunosorbent Assay
    Humans
    Immunoblotting
    Immunoglobulin G
    Immunoglobulin M
    Lyme Disease
    Recombinant Proteins
    Sensitivity and Specificity

    Pub Type(s)

    Evaluation Studies
    Journal Article

    Language

    eng

    PubMed ID

    21271270

    Citation

    TY - JOUR T1 - Large differences between test strategies for the detection of anti-Borrelia antibodies are revealed by comparing eight ELISAs and five immunoblots. AU - Ang,C W, AU - Notermans,D W, AU - Hommes,M, AU - Simoons-Smit,A M, AU - Herremans,T, Y1 - 2011/01/27/ PY - 2010/07/21/received PY - 2011/01/01/accepted PY - 2011/1/29/entrez PY - 2011/1/29/pubmed PY - 2011/10/28/medline SP - 1027 EP - 32 JF - European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology JO - Eur. J. Clin. Microbiol. Infect. Dis. VL - 30 IS - 8 N2 - We investigated the influence of assay choice on the results in a two-tier testing algorithm for the detection of anti-Borrelia antibodies. Eighty-nine serum samples from clinically well-defined patients were tested in eight different enzyme-linked immunosorbent assay (ELISA) systems based on whole-cell antigens, whole-cell antigens supplemented with VlsE and assays using exclusively recombinant proteins. A subset of samples was tested in five immunoblots: one whole-cell blot, one whole-cell blot supplemented with VlsE and three recombinant blots. The number of IgM- and/or IgG-positive ELISA results in the group of patients suspected of Borrelia infection ranged from 34 to 59%. The percentage of positives in cross-reactivity controls ranged from 0 to 38%. Comparison of immunoblots yielded large differences in inter-test agreement and showed, at best, a moderate agreement between tests. Remarkably, some immunoblots gave positive results in samples that had been tested negative by all eight ELISAs. The percentage of positive blots following a positive ELISA result depended heavily on the choice of ELISA-immunoblot combination. We conclude that the assays used to detect anti-Borrelia antibodies have widely divergent sensitivity and specificity. The choice of ELISA-immunoblot combination severely influences the number of positive results, making the exchange of test results between laboratories with different methodologies hazardous. SN - 1435-4373 UR - https://www.unboundmedicine.com/medline/citation/21271270/abstract/Large_differences_between_test_strategies_for_the_detection_of_anti_Borrelia_antibodies_are_revealed_by_comparing_eight_ELISAs_and_five_immunoblots_ L2 - https://dx.doi.org/10.1007/s10096-011-1157-6 ER -