RESEARCH ARTICLE Quantification of Leishmania (Viannia) Kinetoplast DNA in Ulcers of Cutaneous Leishmaniasis Reveals Inter-site and Inter- sampling Variability in Parasite Load

Background Cutaneous leishmaniasis (CL) is a skin disease caused by the protozoan parasite Leish-mania. Few studies have assessed the influence of the sample collection site within the ulcer and the sampling method on the sensitivity of parasitological and molecular diagnostic techniques for CL. Sensitivity of the technique can be dependent upon the load and distribu-tion of Leishmania amastigotes in the lesion. Methodology/Principal Findings We applied a quantitative real-time PCR (qPCR) assay for Leishmania (Viannia) minicircle kinetoplast DNA (kDNA) detection and parasite load quantification in biopsy and scraping samples obtained from 3 sites within each ulcer (border, base, and center) as well as in cytology brush specimens taken from the ulcer base and center. A total of 248 lesion sam-ples from 31 patients with laboratory confirmed CL of recent onset (3 months) were evalu-ated. The kDNA-qPCR detected Leishmania DNA in 97.6 % (242/248) of the examined samples. Median parasite loads were significantly higher in the ulcer base and center tha