Detection of Bartonella quintana DNA in the presence of human and feline whole blood by single-tube PCR without DNA extraction

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.