Title

Enhanced Detection of Host Response Antibodies to Borrelia burgdorferi Using Immuno-PCR

Authors

Authors

M. D. Halpern; S. Jain;M. W. Jewett

Comments

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Abbreviated Journal Title

Clin. Vaccine Immunol.

Keywords

EARLY LYME-DISEASE; DIAGNOSIS; ASSAY; PROTEIN; SERODIAGNOSIS; AMPLIFICATION; ANTIGENS; GENES; TICKS; Immunology; Infectious Diseases; Microbiology

Abstract

Lyme disease is the fastest-growing zoonotic disease in North America. Current methods for detection of Borrelia burgdorferi infection are challenged by analysis subjectivity and standardization of antigen source. In the present study, we developed an immuno-PCR (iPCR)-based approach employing recombinant in vivo-expressed B. burgdorferi antigens for objective detection of a host immune response to B. burgdorferi infection. iPCR is a liquid-phase protein detection method that combines the sensitivity of PCR with the specificity and versatility of immunoassay-based protocols. Use of magnetic beads coated with intact spirochetes provided effective antigen presentation and allowed detection of host-generated antibodies in experimentally infected mice at day 11 postinoculation, whereas host-generated antibodies were detected at day 14 by enzyme-linked immunosorbent assay (ELISA) and day 21 by immunoblotting. Furthermore, magnetic beads coated with recombinant B. burgdorferi in vivo-expressed antigen OspC or BmpA demonstrated positive detection of host-generated antibodies in mice at day 7 postinoculation with markedly increased iPCR signals above the background, with the quantification cycle (C-q) value for each sample minus the mean background C-q plus 3 standard deviations (Delta C-q) being 4 to 10, whereas Delta C-q was 2.5 for intact spirochete-coated beads. iPCR demonstrated a strong correlation (Spearman rank correlation = 0.895, P < 0.0001) with a commercial ELISA for detection of host antibodies in human Lyme disease patient sera using the B. burgdorferi VlsE C6 peptide. In addition, iPCR showed potential applicability for direct detection of spirochetes in blood. The results presented here indicate that our iPCR assay has the potential to provide an objective format that can be used for sensitive detection of multiple host response antibodies and isotypes to B. burgdorferi infection.

Journal Title

Clinical and Vaccine Immunology

Volume

20

Issue/Number

3

Publication Date

1-1-2013

Document Type

Article

Language

English

First Page

350

Last Page

357

WOS Identifier

WOS:000315411100004

ISSN

1556-6811

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