Rapid detection and differentiation of Bartonella spp. by a single-run real-time PCR.

Abstract A Real-time PCR for the identification and differentiation of Bartonella spp. based on the detection of mutations in an internal region of the glt A gene by thermal analysis was developed. The assay included a simultaneous detection of the amplicons by direct hybridization of LCRed and fluorescein-labelled probes coupled with melting curve analysis by the use of fluorescence resonance energy transfer technology. The protocol allowed establishing genospecie identity in less than 1 h without a need for restriction enzyme digestion or gel electrophoresis. The method is simple, reproducible, rapid, specific and potentially transferable to clinical samples.