Identification of Six New World Leishmania species through the implementation of a High-Resolution Melting (HRM) genotyping assay
Background: Leishmaniases are tropical zoonotic diseases, caused by parasites from the genus Leishmania. New World (NW) species are related to sylvatic cycles although urbanization processes have been reported in some South American Countries such as Colombia. This eco-epidemiological complexity imp...
- Autores:
- Tipo de recurso:
- Fecha de publicación:
- 2014
- Institución:
- Universidad del Rosario
- Repositorio:
- Repositorio EdocUR - U. Rosario
- Idioma:
- eng
- OAI Identifier:
- oai:repository.urosario.edu.co:10336/22754
- Acceso en línea:
- https://doi.org/10.1186/s13071-014-0501-y
https://repository.urosario.edu.co/handle/10336/22754
- Palabra clave:
- Heat shock protein 70
Internal transcribed spacer 1
Monoclonal antibody
Protozoal dna
Accuracy
Algorithm
Article
Colombia
Controlled study
Disease carrier
Genotype
High resolution melting analysis
Human
Leishmania
Leishmania amazonensis
Leishmania braziliensis
Leishmania guyanensis
Leishmania infantum
Leishmania mexicana
Leishmania panamensis
Limit of detection
Multilocus enzyme electrophoresis
Nonhuman
Parasite identification
Parasite isolation
Polymerase chain reaction
Reliability
Restriction fragment length polymorphism
Animal
Chemistry
Classification
Evaluation study
Genetic procedures
Genetics
Genotype
Isolation and purification
Leishmania
Leishmaniasis
Mammal
Parasitology
Transition temperature
Animals
Genetic techniques
Genotype
Humans
Insect vectors
Leishmania
Leishmaniasis
Mammals
Transition temperature
Genotyping
High-resolution melting
Leishmania
Real-time pcr
protozoan
Dna
- Rights
- License
- Abierto (Texto Completo)
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dc.title.spa.fl_str_mv |
Identification of Six New World Leishmania species through the implementation of a High-Resolution Melting (HRM) genotyping assay |
title |
Identification of Six New World Leishmania species through the implementation of a High-Resolution Melting (HRM) genotyping assay |
spellingShingle |
Identification of Six New World Leishmania species through the implementation of a High-Resolution Melting (HRM) genotyping assay Heat shock protein 70 Internal transcribed spacer 1 Monoclonal antibody Protozoal dna Accuracy Algorithm Article Colombia Controlled study Disease carrier Genotype High resolution melting analysis Human Leishmania Leishmania amazonensis Leishmania braziliensis Leishmania guyanensis Leishmania infantum Leishmania mexicana Leishmania panamensis Limit of detection Multilocus enzyme electrophoresis Nonhuman Parasite identification Parasite isolation Polymerase chain reaction Reliability Restriction fragment length polymorphism Animal Chemistry Classification Evaluation study Genetic procedures Genetics Genotype Isolation and purification Leishmania Leishmaniasis Mammal Parasitology Transition temperature Animals Genetic techniques Genotype Humans Insect vectors Leishmania Leishmaniasis Mammals Transition temperature Genotyping High-resolution melting Leishmania Real-time pcr protozoan Dna |
title_short |
Identification of Six New World Leishmania species through the implementation of a High-Resolution Melting (HRM) genotyping assay |
title_full |
Identification of Six New World Leishmania species through the implementation of a High-Resolution Melting (HRM) genotyping assay |
title_fullStr |
Identification of Six New World Leishmania species through the implementation of a High-Resolution Melting (HRM) genotyping assay |
title_full_unstemmed |
Identification of Six New World Leishmania species through the implementation of a High-Resolution Melting (HRM) genotyping assay |
title_sort |
Identification of Six New World Leishmania species through the implementation of a High-Resolution Melting (HRM) genotyping assay |
dc.subject.keyword.spa.fl_str_mv |
Heat shock protein 70 Internal transcribed spacer 1 Monoclonal antibody Protozoal dna Accuracy Algorithm Article Colombia Controlled study Disease carrier Genotype High resolution melting analysis Human Leishmania Leishmania amazonensis Leishmania braziliensis Leishmania guyanensis Leishmania infantum Leishmania mexicana Leishmania panamensis Limit of detection Multilocus enzyme electrophoresis Nonhuman Parasite identification Parasite isolation Polymerase chain reaction Reliability Restriction fragment length polymorphism Animal Chemistry Classification Evaluation study Genetic procedures Genetics Genotype Isolation and purification Leishmania Leishmaniasis Mammal Parasitology Transition temperature Animals Genetic techniques Genotype Humans Insect vectors Leishmania Leishmaniasis Mammals Transition temperature Genotyping High-resolution melting Leishmania Real-time pcr |
topic |
Heat shock protein 70 Internal transcribed spacer 1 Monoclonal antibody Protozoal dna Accuracy Algorithm Article Colombia Controlled study Disease carrier Genotype High resolution melting analysis Human Leishmania Leishmania amazonensis Leishmania braziliensis Leishmania guyanensis Leishmania infantum Leishmania mexicana Leishmania panamensis Limit of detection Multilocus enzyme electrophoresis Nonhuman Parasite identification Parasite isolation Polymerase chain reaction Reliability Restriction fragment length polymorphism Animal Chemistry Classification Evaluation study Genetic procedures Genetics Genotype Isolation and purification Leishmania Leishmaniasis Mammal Parasitology Transition temperature Animals Genetic techniques Genotype Humans Insect vectors Leishmania Leishmaniasis Mammals Transition temperature Genotyping High-resolution melting Leishmania Real-time pcr protozoan Dna |
dc.subject.keyword.eng.fl_str_mv |
protozoan Dna |
description |
Background: Leishmaniases are tropical zoonotic diseases, caused by parasites from the genus Leishmania. New World (NW) species are related to sylvatic cycles although urbanization processes have been reported in some South American Countries such as Colombia. This eco-epidemiological complexity imposes a challenge to the detection of circulating parasite species, not only related to human cases but also infecting vectors and reservoirs. Currently, no harmonized methods have been deployed to discriminate the NW Leishmania species. Findings: Herein, we conducted a systematic and mechanistic High-Resolution Melting (HRM) assay targeted to HSP70 and ITS1. Specific primers were designed that coupled with a HRM analyses permitted to discriminate six NW Leishmania species. In order to validate the herein described algorithm, we included 35 natural isolates obtained from human cases, insect vectors and mammals. Our genotyping assay allowed the correct assignment of the six NW Leishmania species (L. mexicana, L. infantum (chagasi), L. amazonensis, L. panamensis, L. guyanensis and L. braziliensis) based on reference strains. When the algorithm was applied to a set of well-characterized strains by means of PCR-RFLP, MLEE and monoclonal antibodies (MA) we observed a tailored concordance between the HRM and PCR-RFLP/MLEE/MA (KI = 1.0). Additionally, we tested the limit of detection for the HRM method showing that this is able to detect at least 10 equivalent-parasites per mL. Conclusions: This is a rapid and reliable method to conduct molecular epidemiology and host-parasite association studies in endemic areas. © 2014 Baleela et al. |
publishDate |
2014 |
dc.date.created.spa.fl_str_mv |
2014 |
dc.date.accessioned.none.fl_str_mv |
2020-05-25T23:57:50Z |
dc.date.available.none.fl_str_mv |
2020-05-25T23:57:50Z |
dc.type.eng.fl_str_mv |
article |
dc.type.coarversion.fl_str_mv |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
dc.type.coar.fl_str_mv |
http://purl.org/coar/resource_type/c_6501 |
dc.type.spa.spa.fl_str_mv |
Artículo |
dc.identifier.doi.none.fl_str_mv |
https://doi.org/10.1186/s13071-014-0501-y |
dc.identifier.issn.none.fl_str_mv |
17563305 |
dc.identifier.uri.none.fl_str_mv |
https://repository.urosario.edu.co/handle/10336/22754 |
url |
https://doi.org/10.1186/s13071-014-0501-y https://repository.urosario.edu.co/handle/10336/22754 |
identifier_str_mv |
17563305 |
dc.language.iso.spa.fl_str_mv |
eng |
language |
eng |
dc.relation.citationIssue.none.fl_str_mv |
No. 1 |
dc.relation.citationTitle.none.fl_str_mv |
Parasites and Vectors |
dc.relation.citationVolume.none.fl_str_mv |
Vol. 7 |
dc.relation.ispartof.spa.fl_str_mv |
Parasites and Vectors, ISSN:17563305, Vol.7, No.1 (2014) |
dc.relation.uri.spa.fl_str_mv |
https://www.scopus.com/inward/record.uri?eid=2-s2.0-84964696967&doi=10.1186%2fs13071-014-0501-y&partnerID=40&md5=f6dfb33f139c4103d421d9d2916ecef4 |
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http://purl.org/coar/access_right/c_abf2 |
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Abierto (Texto Completo) |
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Abierto (Texto Completo) http://purl.org/coar/access_right/c_abf2 |
dc.format.mimetype.none.fl_str_mv |
application/pdf |
dc.publisher.spa.fl_str_mv |
BioMed Central Ltd. |
institution |
Universidad del Rosario |
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instname:Universidad del Rosario |
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reponame:Repositorio Institucional EdocUR |
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5208579560060e26175-15af-4735-87d0-aca34eb413cec6dc2ff7-8da5-4cad-92a4-4fc35b8e82475c82b260-bc7e-4d78-b9a7-97e620c18950bb4882e7-b31a-4fc0-8592-11afa40847da10117161186002020-05-25T23:57:50Z2020-05-25T23:57:50Z2014Background: Leishmaniases are tropical zoonotic diseases, caused by parasites from the genus Leishmania. New World (NW) species are related to sylvatic cycles although urbanization processes have been reported in some South American Countries such as Colombia. This eco-epidemiological complexity imposes a challenge to the detection of circulating parasite species, not only related to human cases but also infecting vectors and reservoirs. Currently, no harmonized methods have been deployed to discriminate the NW Leishmania species. Findings: Herein, we conducted a systematic and mechanistic High-Resolution Melting (HRM) assay targeted to HSP70 and ITS1. Specific primers were designed that coupled with a HRM analyses permitted to discriminate six NW Leishmania species. In order to validate the herein described algorithm, we included 35 natural isolates obtained from human cases, insect vectors and mammals. Our genotyping assay allowed the correct assignment of the six NW Leishmania species (L. mexicana, L. infantum (chagasi), L. amazonensis, L. panamensis, L. guyanensis and L. braziliensis) based on reference strains. When the algorithm was applied to a set of well-characterized strains by means of PCR-RFLP, MLEE and monoclonal antibodies (MA) we observed a tailored concordance between the HRM and PCR-RFLP/MLEE/MA (KI = 1.0). Additionally, we tested the limit of detection for the HRM method showing that this is able to detect at least 10 equivalent-parasites per mL. Conclusions: This is a rapid and reliable method to conduct molecular epidemiology and host-parasite association studies in endemic areas. © 2014 Baleela et al.application/pdfhttps://doi.org/10.1186/s13071-014-0501-y17563305https://repository.urosario.edu.co/handle/10336/22754engBioMed Central Ltd.No. 1Parasites and VectorsVol. 7Parasites and Vectors, ISSN:17563305, Vol.7, No.1 (2014)https://www.scopus.com/inward/record.uri?eid=2-s2.0-84964696967&doi=10.1186%2fs13071-014-0501-y&partnerID=40&md5=f6dfb33f139c4103d421d9d2916ecef4Abierto (Texto Completo)http://purl.org/coar/access_right/c_abf2instname:Universidad del Rosarioreponame:Repositorio Institucional EdocURHeat shock protein 70Internal transcribed spacer 1Monoclonal antibodyProtozoal dnaAccuracyAlgorithmArticleColombiaControlled studyDisease carrierGenotypeHigh resolution melting analysisHumanLeishmaniaLeishmania amazonensisLeishmania braziliensisLeishmania guyanensisLeishmania infantumLeishmania mexicanaLeishmania panamensisLimit of detectionMultilocus enzyme electrophoresisNonhumanParasite identificationParasite isolationPolymerase chain reactionReliabilityRestriction fragment length polymorphismAnimalChemistryClassificationEvaluation studyGenetic proceduresGeneticsGenotypeIsolation and purificationLeishmaniaLeishmaniasisMammalParasitologyTransition temperatureAnimalsGenetic techniquesGenotypeHumansInsect vectorsLeishmaniaLeishmaniasisMammalsTransition temperatureGenotypingHigh-resolution meltingLeishmaniaReal-time pcrprotozoanDnaIdentification of Six New World Leishmania species through the implementation of a High-Resolution Melting (HRM) genotyping assayarticleArtículohttp://purl.org/coar/version/c_970fb48d4fbd8a85http://purl.org/coar/resource_type/c_6501Galindo Hernández, CarolinaAlvarez, CatalinaGonzález, CamilaAyala, Martha StellaLeón, Cielo MaritzaRamírez, Juan DavidORIGINALs13071-014-0501-y.pdfapplication/pdf1040807https://repository.urosario.edu.co/bitstreams/7daae947-88ee-4f4c-84b7-a2ef6a36d3d9/downloadfa17b7d568eda2327210bb479ae876e2MD51TEXTs13071-014-0501-y.pdf.txts13071-014-0501-y.pdf.txtExtracted texttext/plain34059https://repository.urosario.edu.co/bitstreams/b27eb700-c9c9-4198-b37b-29e839a99c4b/download5fa903980d1425217d16bc9a674fa47cMD52THUMBNAILs13071-014-0501-y.pdf.jpgs13071-014-0501-y.pdf.jpgGenerated Thumbnailimage/jpeg4546https://repository.urosario.edu.co/bitstreams/4f4fd9f8-2d4e-481f-92aa-5211b75d5a4c/download91f77a4aa0859a6c6a305e7a11ede690MD5310336/22754oai:repository.urosario.edu.co:10336/227542022-05-02 07:37:13.049115https://repository.urosario.edu.coRepositorio institucional EdocURedocur@urosario.edu.co |