Evaluation of a Novel Microarray Immunoblot Assay for Detection of IgM- and IgG-Class Antibodies to Borrelia burgdorferi

Abstract

Diagnostic testing for Lyme disease (LD) remains dependent on detection of antibodies to LD using serologic assays, in adherence to the standard two-tiered testing (STTT) algorithm. We present the first analytic evaluation of the automated B31 ViraChip IgM and IgG microarray immunoblot (MIB) assays (Viramed Biotech AG, Planegg, Germany) in comparison to two different, semiautomated blot assays for LD, including the B31 ViraStripe IgM and IgG line immunoassays (LIAs) (Viramed) and the MarDx IgM and IgG Western blot (WB) assays (Trinity Biotech, Carlsbad, CA), using prospectively collected sera ( = 411) and archived, clinically characterized samples ( = 91). We show comparable overall agreement (>84%) of the ViraChip MIB assays against the two aforementioned LD blot methods. The ViraChip MIB assays were also compared to a consensus standard, whereby samples were classified as positive or negative for IgM or IgG to if the analyte-matched ViraStripe LIA or MarDx WB assay were positive or negative, respectively. The ViraChip IgM and IgG MIB assays showed>93% positive, negative, and overall agreement versus these consensus criteria. The ViraChip MIB assays were associated with a time savings of 28 min to process one full batch of samples compared to the time required for the ViraStripe LIAs. The ViraChip MIB assays can be programmed and performed on an open-system, automated enzyme-linked immunosorbent assay (ELISA) processor, negating the need for assay-specific equipment and enabling laboratories to consolidate LD testing onto a single platform. We conclude that the ViraChip IgM and IgG MIB assays may be added to the repertoire of supplemental, second-tier blot testing systems for diagnosis of LD.

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

“…In conclusion, several investigations using the multi-antigen approach to improve serologic testing for Lyme disease have been reported underscoring the rationalization for adding antigens for new test algorithms (14–18). A commercial assay would likely employ multiplex testing technology to enhance sensitivity over ELISA formats and provide a platform to screen dozens of antigens (11, 19, 20). These studies and ours represent pilot versions of algorithms for new tests and warrant validation with higher numbers of prospectively and retrospectively collected patient samples.…”

Immunoserological Diagnosis of Human Borrelioses: Current Knowledge and Perspectives

“…In neuroborreliosis from two European countries, the IgG seroreactivity of VlsE alone surpassed that of other antigens commonly used (p100, p58, p39, OspA, OspC, and p18) compared to control patients (Dessau et al, 2015). IB have been recently miniaturized in a microarray format with probable equivalent performances to other commercial immunoblots (Theel et al, 2018). Given the diversity of genospecies involved in Lyme borreliosis in Europe compared to the US, almost all of the available IB kits use combinations of recombinant antigens from strains belonging to different species, generally the three main European pathogenic species (B. afzelii, B. garinii, B. burgdorferi ss) possibly associated with B. bavariensis and B. spielmanii, also responsible for some cases of Lyme borreliosis.…”

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

“…The STTT has been a useful diagnostic tool since its standardization by the Centers for Disease Control (CDC) in 1995 [13], but critical limitations have become increasingly apparent [16,17]. These include low sensitivity and low specificity for early disease [18,19], inability to monitor treatment progress or diagnose re-infection [20], inconsistencies across tests [21][22][23], and subjective interpretation of Western blot results [18,24]. Experts agree that new strategies for diagnosing LD are necessary to address these concerns, pointing to modified two-tier algorithms using only ELISAs [23,[25][26][27], as well as novel assays in various stages of development [28][29][30].…”

“…In this study, we systematically optimized and validated the GC-FP biochip for detection of specific serum antibodies in patients with untreated LD, patients treated for LD with antibiotics, and various negative controls using well characterized serum samples [22,43]. 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.…”