COLORIMETRIC SOLID-PHASE CAPTURE HYBRIDIZATION ASSAY FOR DETECTION OFAMPLIFIED BORRELIA-BURGDORFERI DNA

To simplify detection procedures of DNA fragments resulting from PCR, we developed a colorimetric microplate hybridization assay. This forma t was used for the identification of Borrelia burgdorferi sensu lato, the causal agent of Lyme disease. The system relied on the use of a sp ecific capture probe covalently linked to polystyrene plates and a spe cific polybiotinylated detection probe. DNA fragments, resulting from PCR and sandwiched between these two probes, were detected by enzymati c color development. The new detection format out-performed agarose ge l electrophoresis of PCR products in sensitivity and specificity. More over, in view of its rapidity and simplicity, the system proved approp riate for the routing diagnostic analysis of clinical specimens from L yme disease patients. The proposed detection format can be adapted eas ily to other DNA targets and is suitable for automation.