Simple Objective Detection of Human Lyme Disease Infection Using Immuno-PCR and a Single Recombinant Hybrid Antigen

Clinical and vaccine Immunology: CVI (Impact Factor: 2.47). 06/2014; 21(8). DOI: 10.1128/CVI.00245-14
Source: PubMed


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

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