DSpace Repository

Real-time PCR assay for detection and quantification of Leishmania (Viannia) organisms in skin and mucosal lesions: Exploratory study of parasite load and clinical parameters

Show simple item record

dc.contributor.author Jara, Marlene
dc.contributor.author Adaui, Vanessa
dc.contributor.author Valencia, Braulio M.
dc.contributor.author Martinez, Dalila
dc.contributor.author Alba, Milena
dc.contributor.author Castrillon, Carlos
dc.contributor.author Cruz, Maria
dc.contributor.author Cruz, Israel
dc.contributor.author Van der Auwera, Gert
dc.contributor.author Llanos Cuentas, Elmer Alejandro
dc.contributor.author Dujardin, Jean-Claude
dc.contributor.author Arévalo Zelada, Jorge Luis
dc.date.accessioned 2022-01-04T20:31:42Z
dc.date.available 2022-01-04T20:31:42Z
dc.date.issued 2013
dc.identifier.uri https://hdl.handle.net/20.500.12866/10546
dc.description.abstract Earlier histopathology studies suggest that parasite loads may differ between cutaneous leishmaniasis (CL) and mucosal leishmaniasis (ML) lesions and between acute and chronic CL. Formal demonstration requires highly sensitive detection and accurate quantification of Leishmania in human lesional tissue. In this study, we developed a quantitative real-time PCR (qPCR) assay targeting minicircle kinetoplast DNA (kDNA) to detect and quantify Leishmania (Viannia) parasites. We evaluated a total of 156 lesion biopsy specimens from CL or ML suspected cases and compared the quantitative performance of our kDNA qPCR assay with that of a previously validated qPCR assay based on the glucose-6-phosphate dehydrogenase (G6PD) gene. We also examined the relationship between parasite load and clinical parameters. The kDNA qPCR sensitivity for Leishmania detection was 97.9%, and its specificity was 87.5%. The parasite loads quantified by kDNA qPCR and G6PD qPCR assays were highly correlated (r = 0.87; P < 0.0001), but the former showed higher sensitivity (P = 0.000). CL lesions had 10-fold-higher parasite loads than ML lesions (P = 0.009). Among CL patients, the parasite load was inversely correlated with disease duration (P = 0.004), but there was no difference in parasite load according to the parasite species, the patient's age, and number or area of lesions. Our findings confirm that CL and recent onset of disease (<3 months) are associated with a high parasite load. Our kDNA qPCR assay proved highly sensitive and accurate for the detection and quantification of Leishmania (Viannia) spp. in lesion biopsy specimens. It has potential application as a diagnostic and follow-up tool in American tegumentary leishmaniasis. en_US
dc.language.iso eng
dc.publisher American Society for Microbiology
dc.relation.ispartofseries Journal of Clinical Microbiology
dc.rights info:eu-repo/semantics/restrictedAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
dc.subject Humans en_US
dc.subject Sensitivity and Specificity en_US
dc.subject Real-Time Polymerase Chain Reaction methods en_US
dc.subject DNA, Kinetoplast en_US
dc.subject DNA en_US
dc.subject Leishmania en_US
dc.subject isolation & purification en_US
dc.subject Leishmaniasis en_US
dc.subject Mucous Membrane en_US
dc.subject Parasite Load en_US
dc.subject Skin en_US
dc.title Real-time PCR assay for detection and quantification of Leishmania (Viannia) organisms in skin and mucosal lesions: Exploratory study of parasite load and clinical parameters en_US
dc.type info:eu-repo/semantics/article
dc.identifier.doi https://doi.org/10.1128/JCM.00208-13
dc.subject.ocde https://purl.org/pe-repo/ocde/ford#1.06.01
dc.relation.issn 1098-660X


Files in this item

Files Size Format View

There are no files associated with this item.

This item appears in the following Collection(s)

Show simple item record

info:eu-repo/semantics/restrictedAccess Except where otherwise noted, this item's license is described as info:eu-repo/semantics/restrictedAccess

Search DSpace


Browse

My Account

Statistics