High Resolution Melting Analysis Targeting hsp70 as a Fast and Efficient Method for the Discrimination of Leishmania Species.
BACKGROUND:Protozoan parasites of the genus Leishmania cause a large spectrum of clinical manifestations known as Leishmaniases. These diseases are increasingly important public health problems in many countries both within and outside endemic regions. Thus, an accurate differential diagnosis is ext...
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ftdoajarticles:oai:doaj.org/article:09ae29bc078d422693ce43437aaa2382 2023-05-15T15:09:18+02:00 High Resolution Melting Analysis Targeting hsp70 as a Fast and Efficient Method for the Discrimination of Leishmania Species. Ricardo Andrade Zampieri Maria Fernanda Laranjeira-Silva Sandra Marcia Muxel Ana Carolina Stocco de Lima Jeffrey Jon Shaw Lucile Maria Floeter-Winter 2016-02-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0004485 https://doaj.org/article/09ae29bc078d422693ce43437aaa2382 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC4771719?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0004485 https://doaj.org/article/09ae29bc078d422693ce43437aaa2382 PLoS Neglected Tropical Diseases, Vol 10, Iss 2, p e0004485 (2016) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2016 ftdoajarticles https://doi.org/10.1371/journal.pntd.0004485 2022-12-31T05:36:28Z BACKGROUND:Protozoan parasites of the genus Leishmania cause a large spectrum of clinical manifestations known as Leishmaniases. These diseases are increasingly important public health problems in many countries both within and outside endemic regions. Thus, an accurate differential diagnosis is extremely relevant for understanding epidemiological profiles and for the administration of the best therapeutic protocol. METHODS/PRINCIPAL FINDINGS:Exploring the High Resolution Melting (HRM) dissociation profiles of two amplicons using real time polymerase chain reaction (real-time PCR) targeting heat-shock protein 70 coding gene (hsp70) revealed differences that allowed the discrimination of genomic DNA samples of eight Leishmania species found in the Americas, including Leishmania (Leishmania) infantum chagasi, L. (L.) amazonensis, L. (L.) mexicana, L. (Viannia) lainsoni, L. (V.) braziliensis, L. (V.) guyanensis, L. (V.) naiffi and L. (V.) shawi, and three species found in Eurasia and Africa, including L. (L.) tropica, L. (L.) donovani and L. (L.) major. In addition, we tested DNA samples obtained from standard promastigote culture, naturally infected phlebotomines, experimentally infected mice and clinical human samples to validate the proposed protocol. CONCLUSIONS/SIGNIFICANCE:HRM analysis of hsp70 amplicons is a fast and robust strategy that allowed for the detection and discrimination of all Leishmania species responsible for the Leishmaniases in Brazil and Eurasia/Africa with high sensitivity and accuracy. This method could detect less than one parasite per reaction, even in the presence of host DNA. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 10 2 e0004485 |
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Open Polar |
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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 Ricardo Andrade Zampieri Maria Fernanda Laranjeira-Silva Sandra Marcia Muxel Ana Carolina Stocco de Lima Jeffrey Jon Shaw Lucile Maria Floeter-Winter High Resolution Melting Analysis Targeting hsp70 as a Fast and Efficient Method for the Discrimination of Leishmania Species. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
BACKGROUND:Protozoan parasites of the genus Leishmania cause a large spectrum of clinical manifestations known as Leishmaniases. These diseases are increasingly important public health problems in many countries both within and outside endemic regions. Thus, an accurate differential diagnosis is extremely relevant for understanding epidemiological profiles and for the administration of the best therapeutic protocol. METHODS/PRINCIPAL FINDINGS:Exploring the High Resolution Melting (HRM) dissociation profiles of two amplicons using real time polymerase chain reaction (real-time PCR) targeting heat-shock protein 70 coding gene (hsp70) revealed differences that allowed the discrimination of genomic DNA samples of eight Leishmania species found in the Americas, including Leishmania (Leishmania) infantum chagasi, L. (L.) amazonensis, L. (L.) mexicana, L. (Viannia) lainsoni, L. (V.) braziliensis, L. (V.) guyanensis, L. (V.) naiffi and L. (V.) shawi, and three species found in Eurasia and Africa, including L. (L.) tropica, L. (L.) donovani and L. (L.) major. In addition, we tested DNA samples obtained from standard promastigote culture, naturally infected phlebotomines, experimentally infected mice and clinical human samples to validate the proposed protocol. CONCLUSIONS/SIGNIFICANCE:HRM analysis of hsp70 amplicons is a fast and robust strategy that allowed for the detection and discrimination of all Leishmania species responsible for the Leishmaniases in Brazil and Eurasia/Africa with high sensitivity and accuracy. This method could detect less than one parasite per reaction, even in the presence of host DNA. |
format |
Article in Journal/Newspaper |
author |
Ricardo Andrade Zampieri Maria Fernanda Laranjeira-Silva Sandra Marcia Muxel Ana Carolina Stocco de Lima Jeffrey Jon Shaw Lucile Maria Floeter-Winter |
author_facet |
Ricardo Andrade Zampieri Maria Fernanda Laranjeira-Silva Sandra Marcia Muxel Ana Carolina Stocco de Lima Jeffrey Jon Shaw Lucile Maria Floeter-Winter |
author_sort |
Ricardo Andrade Zampieri |
title |
High Resolution Melting Analysis Targeting hsp70 as a Fast and Efficient Method for the Discrimination of Leishmania Species. |
title_short |
High Resolution Melting Analysis Targeting hsp70 as a Fast and Efficient Method for the Discrimination of Leishmania Species. |
title_full |
High Resolution Melting Analysis Targeting hsp70 as a Fast and Efficient Method for the Discrimination of Leishmania Species. |
title_fullStr |
High Resolution Melting Analysis Targeting hsp70 as a Fast and Efficient Method for the Discrimination of Leishmania Species. |
title_full_unstemmed |
High Resolution Melting Analysis Targeting hsp70 as a Fast and Efficient Method for the Discrimination of Leishmania Species. |
title_sort |
high resolution melting analysis targeting hsp70 as a fast and efficient method for the discrimination of leishmania species. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2016 |
url |
https://doi.org/10.1371/journal.pntd.0004485 https://doaj.org/article/09ae29bc078d422693ce43437aaa2382 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 10, Iss 2, p e0004485 (2016) |
op_relation |
http://europepmc.org/articles/PMC4771719?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0004485 https://doaj.org/article/09ae29bc078d422693ce43437aaa2382 |
op_doi |
https://doi.org/10.1371/journal.pntd.0004485 |
container_title |
PLOS Neglected Tropical Diseases |
container_volume |
10 |
container_issue |
2 |
container_start_page |
e0004485 |
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1766340513889255424 |