A FRET-based real-time PCR assay to identify the main causal agents of New World tegumentary leishmaniasis.
In South America, various species of Leishmania are endemic and cause New World tegumentary leishmaniasis (NWTL). The correct identification of these species is critical for adequate clinical management and surveillance activities. We developed a real-time polymerase chain reaction (PCR) assay and e...
Published in: | PLoS Neglected Tropical Diseases |
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ftdoajarticles:oai:doaj.org/article:e9c2d3adbfd1471cb06018deee281060 2023-05-15T15:11:30+02:00 A FRET-based real-time PCR assay to identify the main causal agents of New World tegumentary leishmaniasis. Pablo Tsukayama Jorge H Núñez Maxy De Los Santos Valeria Soberón Carmen M Lucas Greg Matlashewski Alejandro Llanos-Cuentas Marianela Ore G Christian Baldeviano Kimberly A Edgel Andres G Lescano Paul C F Graf David J Bacon 2013-01-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0001956 https://doaj.org/article/e9c2d3adbfd1471cb06018deee281060 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC3536805?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0001956 https://doaj.org/article/e9c2d3adbfd1471cb06018deee281060 PLoS Neglected Tropical Diseases, Vol 7, Iss 1, p e1956 (2013) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2013 ftdoajarticles https://doi.org/10.1371/journal.pntd.0001956 2022-12-31T00:25:31Z In South America, various species of Leishmania are endemic and cause New World tegumentary leishmaniasis (NWTL). The correct identification of these species is critical for adequate clinical management and surveillance activities. We developed a real-time polymerase chain reaction (PCR) assay and evaluated its diagnostic performance using 64 archived parasite isolates and 192 prospectively identified samples collected from individuals with suspected leishmaniasis enrolled at two reference clinics in Lima, Peru. The real-time PCR assay was able to detect a single parasite and provided unambiguous melting peaks for five Leishmania species of the Viannia subgenus that are highly prevalent in South America: L. (V.) braziliensis, L. (V.) panamensis, L. (V.) guyanensis, L. (V.) peruviana and L. (V.) lainsoni. Using kinetoplastid DNA-based PCR as a gold standard, the real-time PCR had sensitivity and specificity values of 92% and 77%, respectively, which were significantly higher than those of conventional tests such as microscopy, culture and the leishmanin skin test (LST). In addition, the real-time PCR identified 147 different clinical samples at the species level, providing an overall agreement of 100% when compared to multilocus sequence typing (MLST) data performed on a subset of these samples. Furthermore, the real-time PCR was three times faster and five times less expensive when compared to PCR - MLST for species identification from clinical specimens. In summary, this new assay represents a cost-effective and reliable alternative for the identification of the main species causing NWTL in South America. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLoS Neglected Tropical Diseases 7 1 e1956 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
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 Pablo Tsukayama Jorge H Núñez Maxy De Los Santos Valeria Soberón Carmen M Lucas Greg Matlashewski Alejandro Llanos-Cuentas Marianela Ore G Christian Baldeviano Kimberly A Edgel Andres G Lescano Paul C F Graf David J Bacon A FRET-based real-time PCR assay to identify the main causal agents of New World tegumentary leishmaniasis. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
In South America, various species of Leishmania are endemic and cause New World tegumentary leishmaniasis (NWTL). The correct identification of these species is critical for adequate clinical management and surveillance activities. We developed a real-time polymerase chain reaction (PCR) assay and evaluated its diagnostic performance using 64 archived parasite isolates and 192 prospectively identified samples collected from individuals with suspected leishmaniasis enrolled at two reference clinics in Lima, Peru. The real-time PCR assay was able to detect a single parasite and provided unambiguous melting peaks for five Leishmania species of the Viannia subgenus that are highly prevalent in South America: L. (V.) braziliensis, L. (V.) panamensis, L. (V.) guyanensis, L. (V.) peruviana and L. (V.) lainsoni. Using kinetoplastid DNA-based PCR as a gold standard, the real-time PCR had sensitivity and specificity values of 92% and 77%, respectively, which were significantly higher than those of conventional tests such as microscopy, culture and the leishmanin skin test (LST). In addition, the real-time PCR identified 147 different clinical samples at the species level, providing an overall agreement of 100% when compared to multilocus sequence typing (MLST) data performed on a subset of these samples. Furthermore, the real-time PCR was three times faster and five times less expensive when compared to PCR - MLST for species identification from clinical specimens. In summary, this new assay represents a cost-effective and reliable alternative for the identification of the main species causing NWTL in South America. |
format |
Article in Journal/Newspaper |
author |
Pablo Tsukayama Jorge H Núñez Maxy De Los Santos Valeria Soberón Carmen M Lucas Greg Matlashewski Alejandro Llanos-Cuentas Marianela Ore G Christian Baldeviano Kimberly A Edgel Andres G Lescano Paul C F Graf David J Bacon |
author_facet |
Pablo Tsukayama Jorge H Núñez Maxy De Los Santos Valeria Soberón Carmen M Lucas Greg Matlashewski Alejandro Llanos-Cuentas Marianela Ore G Christian Baldeviano Kimberly A Edgel Andres G Lescano Paul C F Graf David J Bacon |
author_sort |
Pablo Tsukayama |
title |
A FRET-based real-time PCR assay to identify the main causal agents of New World tegumentary leishmaniasis. |
title_short |
A FRET-based real-time PCR assay to identify the main causal agents of New World tegumentary leishmaniasis. |
title_full |
A FRET-based real-time PCR assay to identify the main causal agents of New World tegumentary leishmaniasis. |
title_fullStr |
A FRET-based real-time PCR assay to identify the main causal agents of New World tegumentary leishmaniasis. |
title_full_unstemmed |
A FRET-based real-time PCR assay to identify the main causal agents of New World tegumentary leishmaniasis. |
title_sort |
fret-based real-time pcr assay to identify the main causal agents of new world tegumentary leishmaniasis. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2013 |
url |
https://doi.org/10.1371/journal.pntd.0001956 https://doaj.org/article/e9c2d3adbfd1471cb06018deee281060 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 7, Iss 1, p e1956 (2013) |
op_relation |
http://europepmc.org/articles/PMC3536805?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0001956 https://doaj.org/article/e9c2d3adbfd1471cb06018deee281060 |
op_doi |
https://doi.org/10.1371/journal.pntd.0001956 |
container_title |
PLoS Neglected Tropical Diseases |
container_volume |
7 |
container_issue |
1 |
container_start_page |
e1956 |
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1766342343664861184 |