Quantification of Leishmania ( Viannia ) Kinetoplast DNA in Ulcers of Cutaneous Leishmaniasis Reveals Inter-site and Inter-sampling Variability in Parasite Load: e0003936

Background Cutaneous leishmaniasis (CL) is a skin disease caused by the protozoan parasite Leishmania. 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. Sens...

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Bibliographic Details
Published in:PLoS neglected tropical diseases 2015-07, Vol.9 (7)
Main Authors: Suárez, Milagros, Valencia, Braulio M, Jara, Marlene, Alba, Milena, Boggild, Andrea K, Dujardin, Jean-Claude, Llanos-Cuentas, Alejandro, Arevalo, Jorge, Adaui, Vanessa
Format: Article
Language:English
Subjects:
Biopsy
Cellular biology
Confidence intervals
Cooperation
Cytology
Deoxyribonucleic acid
DNA
Epidemiology
Hypothesis testing
Infections
Lesions
Load
Load distribution
Methods
Parasites
Parasitic diseases
Skin
Skin diseases
Studies
Tropical diseases
Vector-borne diseases
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Summary:Background Cutaneous leishmaniasis (CL) is a skin disease caused by the protozoan parasite Leishmania. 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 distribution 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 samples from 31 patients with laboratory confirmed CL of recent onset (≤3 months) were evaluated. 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 than in the border in biopsies (P
ISSN:1935-2727
1935-2735
DOI:10.1371/journal.pntd.0003936