Abstract

Background: Rapid and cost-effective detection of emerging zoonotic blood-borne pathogens, such as Bartonella spp., is important for diagnostic and surveillance purposes. DNA extraction is a tedious process that increases the risk of cross-contamination, decreases the amount of target nucleic acids, and increases costs. The concept of using a direct PCR protocol for the detection of Bartonella in the presence of whole feline or human blood without DNA extraction was evaluated.

Findings: An optimized, single-tube, direct PCR for the detection of B. quintana in the presence of 20% of EDTA whole blood was optimized and validated using a DNA polymerase resistant to common PCR inhibitors. Ten genome equivalents of B. quintana/µl of blood were detected 100% of the time, and 2 genome equivalents/µl were detected 91% of the time.

Conclusions: The direct PCR for the detection of Bartonella quintana in whole blood is a viable, quick, highly sensitive, and cost-effective alternative that deserves further exploration.

Keywords: 16S-23S ribosomal RNA intergenic transcribed spacer, Alphaproteobacteria, Bartonellosis, Diagnosis, Direct PCR