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...

Full description

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)
id EDOCUR2_829b38a2637348ac605a718ed07a74d4
oai_identifier_str oai:repository.urosario.edu.co:10336/22754
network_acronym_str EDOCUR2
network_name_str Repositorio EdocUR - U. Rosario
repository_id_str
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
dc.rights.coar.fl_str_mv http://purl.org/coar/access_right/c_abf2
dc.rights.acceso.spa.fl_str_mv Abierto (Texto Completo)
rights_invalid_str_mv 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
dc.source.instname.spa.fl_str_mv instname:Universidad del Rosario
dc.source.reponame.spa.fl_str_mv reponame:Repositorio Institucional EdocUR
bitstream.url.fl_str_mv https://repository.urosario.edu.co/bitstreams/7daae947-88ee-4f4c-84b7-a2ef6a36d3d9/download
https://repository.urosario.edu.co/bitstreams/b27eb700-c9c9-4198-b37b-29e839a99c4b/download
https://repository.urosario.edu.co/bitstreams/4f4fd9f8-2d4e-481f-92aa-5211b75d5a4c/download
bitstream.checksum.fl_str_mv fa17b7d568eda2327210bb479ae876e2
5fa903980d1425217d16bc9a674fa47c
91f77a4aa0859a6c6a305e7a11ede690
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
MD5
repository.name.fl_str_mv Repositorio institucional EdocUR
repository.mail.fl_str_mv edocur@urosario.edu.co
_version_ 1814167437482917888
spelling 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