Viability and burden of Leishmania in extralesional sites during human dermal leishmaniasis.
The clinical and epidemiological significance of Leishmania DNA in extralesional sites is obscured by uncertainty of whether the DNA derives from viable parasites. To examine dissemination of Leishmania during active disease and the potential participation of human infection in transmission, Leishma...
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ftdoajarticles:oai:doaj.org/article:96ed236699064d7ead3209572f2aae0d 2023-05-15T15:10:39+02:00 Viability and burden of Leishmania in extralesional sites during human dermal leishmaniasis. Ibeth Romero Jair Téllez Yazmín Suárez Maria Cardona Roger Figueroa Adrian Zelazny Nancy Gore Saravia 2010-09-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0000819 https://doaj.org/article/96ed236699064d7ead3209572f2aae0d EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC2939031?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0000819 https://doaj.org/article/96ed236699064d7ead3209572f2aae0d PLoS Neglected Tropical Diseases, Vol 4, Iss 9 (2010) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2010 ftdoajarticles https://doi.org/10.1371/journal.pntd.0000819 2022-12-31T15:58:41Z The clinical and epidemiological significance of Leishmania DNA in extralesional sites is obscured by uncertainty of whether the DNA derives from viable parasites. To examine dissemination of Leishmania during active disease and the potential participation of human infection in transmission, Leishmania 7SLRNA was exploited to establish viability and estimate parasite burden in extralesional sites of dermal leishmaniasis patients.The feasibility of discriminating parasite viability by PCR of Leishmania 7SLRNA was evaluated in relation with luciferase activity of luc transfected intracellular amastigotes in dose-response assays of Glucantime cytotoxicity. Monocytes, tonsil swabs, aspirates of normal skin and lesions of 28 cutaneous and 2 mucocutaneous leishmaniasis patients were screened by kDNA amplification/Southern blot. Positive samples were analyzed by quantitative PCR of Leishmania 7SLRNA genes and transcripts.7SLRNA amplification coincided with luciferase activity, confirming discrimination of parasite viability. Of 22 patients presenting kDNA in extralesional samples, Leishmania 7SLRNA genes or transcripts were detected in one or more kDNA positive samples in 100% and 73% of patients, respectively. Gene and transcript copy number amplified from extralesional tissues were comparable to lesions. 7SLRNA transcripts were detected in 13/19 (68%) monocyte samples, 5/12 (42%) tonsil swabs, 4/11 (36%) normal skin aspirates, and 22/25 (88%) lesions; genes were quantifiable in 15/19 (79%) monocyte samples, 12/13 (92%) tonsil swabs, 8/11 (73%) normal skin aspirates.Viable parasites are present in extralesional sites, including blood monocytes, tonsils and normal skin of dermal leishmaniasis patients. Leishmania 7SLRNA is an informative target for clinical and epidemiologic investigations of human leishmaniasis. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLoS Neglected Tropical Diseases 4 9 e819 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Ibeth Romero Jair Téllez Yazmín Suárez Maria Cardona Roger Figueroa Adrian Zelazny Nancy Gore Saravia Viability and burden of Leishmania in extralesional sites during human dermal leishmaniasis. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
The clinical and epidemiological significance of Leishmania DNA in extralesional sites is obscured by uncertainty of whether the DNA derives from viable parasites. To examine dissemination of Leishmania during active disease and the potential participation of human infection in transmission, Leishmania 7SLRNA was exploited to establish viability and estimate parasite burden in extralesional sites of dermal leishmaniasis patients.The feasibility of discriminating parasite viability by PCR of Leishmania 7SLRNA was evaluated in relation with luciferase activity of luc transfected intracellular amastigotes in dose-response assays of Glucantime cytotoxicity. Monocytes, tonsil swabs, aspirates of normal skin and lesions of 28 cutaneous and 2 mucocutaneous leishmaniasis patients were screened by kDNA amplification/Southern blot. Positive samples were analyzed by quantitative PCR of Leishmania 7SLRNA genes and transcripts.7SLRNA amplification coincided with luciferase activity, confirming discrimination of parasite viability. Of 22 patients presenting kDNA in extralesional samples, Leishmania 7SLRNA genes or transcripts were detected in one or more kDNA positive samples in 100% and 73% of patients, respectively. Gene and transcript copy number amplified from extralesional tissues were comparable to lesions. 7SLRNA transcripts were detected in 13/19 (68%) monocyte samples, 5/12 (42%) tonsil swabs, 4/11 (36%) normal skin aspirates, and 22/25 (88%) lesions; genes were quantifiable in 15/19 (79%) monocyte samples, 12/13 (92%) tonsil swabs, 8/11 (73%) normal skin aspirates.Viable parasites are present in extralesional sites, including blood monocytes, tonsils and normal skin of dermal leishmaniasis patients. Leishmania 7SLRNA is an informative target for clinical and epidemiologic investigations of human leishmaniasis. |
format |
Article in Journal/Newspaper |
author |
Ibeth Romero Jair Téllez Yazmín Suárez Maria Cardona Roger Figueroa Adrian Zelazny Nancy Gore Saravia |
author_facet |
Ibeth Romero Jair Téllez Yazmín Suárez Maria Cardona Roger Figueroa Adrian Zelazny Nancy Gore Saravia |
author_sort |
Ibeth Romero |
title |
Viability and burden of Leishmania in extralesional sites during human dermal leishmaniasis. |
title_short |
Viability and burden of Leishmania in extralesional sites during human dermal leishmaniasis. |
title_full |
Viability and burden of Leishmania in extralesional sites during human dermal leishmaniasis. |
title_fullStr |
Viability and burden of Leishmania in extralesional sites during human dermal leishmaniasis. |
title_full_unstemmed |
Viability and burden of Leishmania in extralesional sites during human dermal leishmaniasis. |
title_sort |
viability and burden of leishmania in extralesional sites during human dermal leishmaniasis. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2010 |
url |
https://doi.org/10.1371/journal.pntd.0000819 https://doaj.org/article/96ed236699064d7ead3209572f2aae0d |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 4, Iss 9 (2010) |
op_relation |
http://europepmc.org/articles/PMC2939031?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0000819 https://doaj.org/article/96ed236699064d7ead3209572f2aae0d |
op_doi |
https://doi.org/10.1371/journal.pntd.0000819 |
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PLoS Neglected Tropical Diseases |
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4 |
container_issue |
9 |
container_start_page |
e819 |
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