Detection of Leishmania RNA virus in Leishmania parasites.
Patients suffering from cutaneous leishmaniasis (CL) caused by New World Leishmania (Viannia) species are at high risk of developing mucosal (ML) or disseminated cutaneous leishmaniasis (DCL). After the formation of a primary skin lesion at the site of the bite by a Leishmania-infected sand fly, the...
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ftdoajarticles:oai:doaj.org/article:982bcc92c4d8457f867d1b6c89236f10 2023-05-15T15:14:24+02:00 Detection of Leishmania RNA virus in Leishmania parasites. Haroun Zangger Catherine Ronet Chantal Desponds F Matthew Kuhlmann John Robinson Mary-Anne Hartley Florence Prevel Patrik Castiglioni Francine Pratlong Patrick Bastien Norbert Müller Laurent Parmentier Nancy Gore Saravia Stephen M Beverley Nicolas Fasel 2013-01-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0002006 https://doaj.org/article/982bcc92c4d8457f867d1b6c89236f10 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC3542153?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0002006 https://doaj.org/article/982bcc92c4d8457f867d1b6c89236f10 PLoS Neglected Tropical Diseases, Vol 7, Iss 1, p e2006 (2013) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2013 ftdoajarticles https://doi.org/10.1371/journal.pntd.0002006 2022-12-31T13:04:06Z Patients suffering from cutaneous leishmaniasis (CL) caused by New World Leishmania (Viannia) species are at high risk of developing mucosal (ML) or disseminated cutaneous leishmaniasis (DCL). After the formation of a primary skin lesion at the site of the bite by a Leishmania-infected sand fly, the infection can disseminate to form secondary lesions. This metastatic phenotype causes significant morbidity and is often associated with a hyper-inflammatory immune response leading to the destruction of nasopharyngeal tissues in ML, and appearance of nodules or numerous ulcerated skin lesions in DCL. Recently, we connected this aggressive phenotype to the presence of Leishmania RNA virus (LRV) in strains of L. guyanensis, showing that LRV is responsible for elevated parasitaemia, destructive hyper-inflammation and an overall exacerbation of the disease. Further studies of this relationship and the distribution of LRVs in other Leishmania strains and species would benefit from improved methods of viral detection and quantitation, especially ones not dependent on prior knowledge of the viral sequence as LRVs show significant evolutionary divergence.This study reports various techniques, among which, the use of an anti-dsRNA monoclonal antibody (J2) stands out for its specific and quantitative recognition of dsRNA in a sequence-independent fashion. Applications of J2 include immunofluorescence, ELISA and dot blot: techniques complementing an arsenal of other detection tools, such as nucleic acid purification and quantitative real-time-PCR. We evaluate each method as well as demonstrate a successful LRV detection by the J2 antibody in several parasite strains, a freshly isolated patient sample and lesion biopsies of infected mice.We propose that refinements of these methods could be transferred to the field for use as a diagnostic tool in detecting the presence of LRV, and potentially assessing the LRV-related risk of complications in cutaneous leishmaniasis. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLoS Neglected Tropical Diseases 7 1 e2006 |
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 |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Haroun Zangger Catherine Ronet Chantal Desponds F Matthew Kuhlmann John Robinson Mary-Anne Hartley Florence Prevel Patrik Castiglioni Francine Pratlong Patrick Bastien Norbert Müller Laurent Parmentier Nancy Gore Saravia Stephen M Beverley Nicolas Fasel Detection of Leishmania RNA virus in Leishmania parasites. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Patients suffering from cutaneous leishmaniasis (CL) caused by New World Leishmania (Viannia) species are at high risk of developing mucosal (ML) or disseminated cutaneous leishmaniasis (DCL). After the formation of a primary skin lesion at the site of the bite by a Leishmania-infected sand fly, the infection can disseminate to form secondary lesions. This metastatic phenotype causes significant morbidity and is often associated with a hyper-inflammatory immune response leading to the destruction of nasopharyngeal tissues in ML, and appearance of nodules or numerous ulcerated skin lesions in DCL. Recently, we connected this aggressive phenotype to the presence of Leishmania RNA virus (LRV) in strains of L. guyanensis, showing that LRV is responsible for elevated parasitaemia, destructive hyper-inflammation and an overall exacerbation of the disease. Further studies of this relationship and the distribution of LRVs in other Leishmania strains and species would benefit from improved methods of viral detection and quantitation, especially ones not dependent on prior knowledge of the viral sequence as LRVs show significant evolutionary divergence.This study reports various techniques, among which, the use of an anti-dsRNA monoclonal antibody (J2) stands out for its specific and quantitative recognition of dsRNA in a sequence-independent fashion. Applications of J2 include immunofluorescence, ELISA and dot blot: techniques complementing an arsenal of other detection tools, such as nucleic acid purification and quantitative real-time-PCR. We evaluate each method as well as demonstrate a successful LRV detection by the J2 antibody in several parasite strains, a freshly isolated patient sample and lesion biopsies of infected mice.We propose that refinements of these methods could be transferred to the field for use as a diagnostic tool in detecting the presence of LRV, and potentially assessing the LRV-related risk of complications in cutaneous leishmaniasis. |
format |
Article in Journal/Newspaper |
author |
Haroun Zangger Catherine Ronet Chantal Desponds F Matthew Kuhlmann John Robinson Mary-Anne Hartley Florence Prevel Patrik Castiglioni Francine Pratlong Patrick Bastien Norbert Müller Laurent Parmentier Nancy Gore Saravia Stephen M Beverley Nicolas Fasel |
author_facet |
Haroun Zangger Catherine Ronet Chantal Desponds F Matthew Kuhlmann John Robinson Mary-Anne Hartley Florence Prevel Patrik Castiglioni Francine Pratlong Patrick Bastien Norbert Müller Laurent Parmentier Nancy Gore Saravia Stephen M Beverley Nicolas Fasel |
author_sort |
Haroun Zangger |
title |
Detection of Leishmania RNA virus in Leishmania parasites. |
title_short |
Detection of Leishmania RNA virus in Leishmania parasites. |
title_full |
Detection of Leishmania RNA virus in Leishmania parasites. |
title_fullStr |
Detection of Leishmania RNA virus in Leishmania parasites. |
title_full_unstemmed |
Detection of Leishmania RNA virus in Leishmania parasites. |
title_sort |
detection of leishmania rna virus in leishmania parasites. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2013 |
url |
https://doi.org/10.1371/journal.pntd.0002006 https://doaj.org/article/982bcc92c4d8457f867d1b6c89236f10 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 7, Iss 1, p e2006 (2013) |
op_relation |
http://europepmc.org/articles/PMC3542153?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0002006 https://doaj.org/article/982bcc92c4d8457f867d1b6c89236f10 |
op_doi |
https://doi.org/10.1371/journal.pntd.0002006 |
container_title |
PLoS Neglected Tropical Diseases |
container_volume |
7 |
container_issue |
1 |
container_start_page |
e2006 |
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1766344861331488768 |