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Simple objective detection of human lyme disease infection using immuno-PCR and a single recombinant hybrid antigen.

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Abstract

A serology-based tiered approach has, to date, provided the most effective means of laboratory confirmation of clinically suspected cases of
Lyme disease, but it lacks sensitivity in the early stages of
disease and is often dependent on subjectively scored immunoblots. We recently demonstrated the use of immuno-PCR (iPCR) for detecting Borrelia burgdorferi antibodies in patient serum samples that were positive for
Lyme disease. To better understand the performance of the
Lyme disease iPCR assay, the repeatability and variability of the background of the assay across samples from a healthy population (n = 36) were analyzed. Both of these parameters were found to have coefficients of variation of <3%. Using eight antigen-specific iPCR assays and positive call thresholds established for each assay, iPCR IgM and/or IgG diagnosis from
Lyme disease patient serum samples (n = 12) demonstrated a strong correlation with that of 2-tier testing. Furthermore, a simplified iPCR approach using a single hybrid antigen and detecting only IgG antibodies confirmed the 2-tier diagnosis in the
Lyme disease patient serum samples (n = 12). Validation of the hybrid antigen IgG iPCR assay using a blinded panel of
Lyme disease and non-
Lyme disease patient serum samples (n = 92) resulted in a sensitivity of 69% (95% confidence interval [CI], 50% to 84%), compared to that of the 2-tier analysis at 59% (95% CI, 41% to 76%), and a specificity of 98% (95% CI, 91% to 100%) compared to that of the 2-tier analysis at 97% (95% CI, 88% to 100%). A single-tier hybrid antigen iPCR assay has the potential to be an improved method for detecting host-generated antibodies against B. burgdorferi.

Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Clin Vaccine Immunol. 2014 Aug;21(8):1094-105. doi: 10.1128/CVI.00245-14. Epub 2014 Jun 4. Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov’t

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