Quantification of Leishmania (Viannia) Kinetoplast DNA in Ulcers of Cutaneous Leishmaniasis Reveals Inter-site and Inter-sampling Variability in Parasite Load
| Autor: | Milena Alba, Braulio M. Valencia, Jorge Arevalo, Vanessa Adaui, Andrea K. Boggild, Alejandro Llanos-Cuentas, Marlene Jara, Jean-Claude Dujardin, Milagros Suarez |
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| Rok vydání: | 2015 |
| Předmět: |
Adult
Male Pathology medicine.medical_specialty Skin Ulcer/parasitology/pathology lcsh:Arctic medicine. Tropical medicine Adolescent Leishmaniasis Cutaneous/parasitology/pathology lcsh:RC955-962 Leishmaniasis Cutaneous DNA Kinetoplast/genetics Parasite load Parasite Load Young Adult Species Specificity Cutaneous leishmaniasis Cytology Leishmania/classification/genetics Skin Ulcer parasitic diseases medicine Humans Parasite hosting Aged Leishmania biology DNA Kinetoplast lcsh:Public aspects of medicine Public Health Environmental and Occupational Health lcsh:RA1-1270 Leishmaniasis Middle Aged Skin ulcer medicine.disease biology.organism_classification Infectious Diseases Female Human medicine Sample collection medicine.symptom purl.org/pe-repo/ocde/ford#3.03.06 [https] Research Article |
| Zdroj: | PLoS neglected tropical diseases PLoS Neglected Tropical Diseases, Vol 9, Iss 7, p e0003936 (2015) PLoS Neglected Tropical Diseases |
| ISSN: | 1935-2735 1935-2727 |
| Popis: | 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 Author Summary Cutaneous leishmaniasis (CL) is a parasitic disease of the skin caused by obligate intra-macrophage protozoa of the genus Leishmania which usually presents as ulcerative lesions at the site of infection. Traditionally, histopathological and diagnostic studies on CL have employed samples collected from the border of the ulcer since this area is believed to contain the highest amount of parasites. However, no formal demonstration of the distribution of Leishmania parasites in the ulcer has been provided yet. Focusing on human skin lesions of recent onset (≤3 months) caused by L. (Viannia) species, we estimated the parasite loads among different skin lesion sites by means of quantitative real-time PCR targeting the parasite kinetoplast DNA. Paired lesion samples collected by use of different sampling methods were analyzed. We found that the ulcerated zone of the lesion contained a higher parasite load than the ulcer border, and that scraping and cytology brush specimens presented higher parasite loads as compared to the more invasive biopsy. Our results have implications for bedside collection of diagnostic and post-therapeutic follow-up specimens from CL patients. |
| Databáze: | OpenAIRE |
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