Detection and identification of old world Leishmania by high resolution melt analysis.

<h4>Background</h4>Three major forms of human disease, cutaneous leishmaniasis, visceral leishmaniasis and mucocutaneous leishmaniasis, are caused by several leishmanial species whose geographic distribution frequently overlaps. These Leishmania species have diverse reservoir hosts, sand...

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Autores principales:	Dalit Talmi-Frank, Abedelmajeed Nasereddin, Lionel F Schnur, Gabriele Schönian, Seray Ozensoy Töz, Charles L Jaffe, Gad Baneth
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2010
Materias:	Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270
Acceso en línea:	https://doaj.org/article/d28273b327594cd5a6d53519c25d75cb
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spelling	oai:doaj.org-article:d28273b327594cd5a6d53519c25d75cb2021-11-25T06:33:43ZDetection and identification of old world Leishmania by high resolution melt analysis.1935-27271935-273510.1371/journal.pntd.0000581https://doaj.org/article/d28273b327594cd5a6d53519c25d75cb2010-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/20069036/?tool=EBIhttps://doaj.org/toc/1935-2727https://doaj.org/toc/1935-2735<h4>Background</h4>Three major forms of human disease, cutaneous leishmaniasis, visceral leishmaniasis and mucocutaneous leishmaniasis, are caused by several leishmanial species whose geographic distribution frequently overlaps. These Leishmania species have diverse reservoir hosts, sand fly vectors and transmission patterns. In the Old World, the main parasite species responsible for leishmaniasis are Leishmania infantum, L. donovani, L. tropica, L. aethiopica and L. major. Accurate, rapid and sensitive diagnostic and identification procedures are crucial for the detection of infection and characterization of the causative leishmanial species, in order to provide accurate treatment, precise prognosis and appropriate public health control measures.<h4>Methods/principal findings</h4>High resolution melt analysis of a real time PCR product from the Internal Transcribed Spacer-1 rRNA region was used to identify and quantify Old World Leishmania in 300 samples from human patients, reservoir hosts and sand flies. Different characteristic high resolution melt analysis patterns were exhibited by L. major, L. tropica, L. aethiopica, and L. infantum. Genotyping by high resolution melt analysis was verified by DNA sequencing or restriction fragment length polymorphism. This new assay was able to detect as little as 2-4 ITS1 gene copies in a 5 microl DNA sample, i.e., less than a single parasite per reaction.<h4>Conclusions/significance</h4>This new technique is useful for rapid diagnosis of leishmaniasis and simultaneous identification and quantification of the infecting Leishmania species. It can be used for diagnostic purposes directly from clinical samples, as well as epidemiological studies, reservoir host investigations and vector surveys.Dalit Talmi-FrankAbedelmajeed NasereddinLionel F SchnurGabriele SchönianSeray Ozensoy TözCharles L JaffeGad BanethPublic Library of Science (PLoS)articleArctic medicine. Tropical medicineRC955-962Public aspects of medicineRA1-1270ENPLoS Neglected Tropical Diseases, Vol 4, Iss 1, p e581 (2010)
institution	DOAJ
collection	DOAJ
language	EN
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 Dalit Talmi-Frank Abedelmajeed Nasereddin Lionel F Schnur Gabriele Schönian Seray Ozensoy Töz Charles L Jaffe Gad Baneth Detection and identification of old world Leishmania by high resolution melt analysis.
description	<h4>Background</h4>Three major forms of human disease, cutaneous leishmaniasis, visceral leishmaniasis and mucocutaneous leishmaniasis, are caused by several leishmanial species whose geographic distribution frequently overlaps. These Leishmania species have diverse reservoir hosts, sand fly vectors and transmission patterns. In the Old World, the main parasite species responsible for leishmaniasis are Leishmania infantum, L. donovani, L. tropica, L. aethiopica and L. major. Accurate, rapid and sensitive diagnostic and identification procedures are crucial for the detection of infection and characterization of the causative leishmanial species, in order to provide accurate treatment, precise prognosis and appropriate public health control measures.<h4>Methods/principal findings</h4>High resolution melt analysis of a real time PCR product from the Internal Transcribed Spacer-1 rRNA region was used to identify and quantify Old World Leishmania in 300 samples from human patients, reservoir hosts and sand flies. Different characteristic high resolution melt analysis patterns were exhibited by L. major, L. tropica, L. aethiopica, and L. infantum. Genotyping by high resolution melt analysis was verified by DNA sequencing or restriction fragment length polymorphism. This new assay was able to detect as little as 2-4 ITS1 gene copies in a 5 microl DNA sample, i.e., less than a single parasite per reaction.<h4>Conclusions/significance</h4>This new technique is useful for rapid diagnosis of leishmaniasis and simultaneous identification and quantification of the infecting Leishmania species. It can be used for diagnostic purposes directly from clinical samples, as well as epidemiological studies, reservoir host investigations and vector surveys.
format	article
author	Dalit Talmi-Frank Abedelmajeed Nasereddin Lionel F Schnur Gabriele Schönian Seray Ozensoy Töz Charles L Jaffe Gad Baneth
author_facet	Dalit Talmi-Frank Abedelmajeed Nasereddin Lionel F Schnur Gabriele Schönian Seray Ozensoy Töz Charles L Jaffe Gad Baneth
author_sort	Dalit Talmi-Frank
title	Detection and identification of old world Leishmania by high resolution melt analysis.
title_short	Detection and identification of old world Leishmania by high resolution melt analysis.
title_full	Detection and identification of old world Leishmania by high resolution melt analysis.
title_fullStr	Detection and identification of old world Leishmania by high resolution melt analysis.
title_full_unstemmed	Detection and identification of old world Leishmania by high resolution melt analysis.
title_sort	detection and identification of old world leishmania by high resolution melt analysis.
publisher	Public Library of Science (PLoS)
publishDate	2010
url	https://doaj.org/article/d28273b327594cd5a6d53519c25d75cb
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Detection and identification of old world Leishmania by high resolution melt analysis.

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