Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identificatio nof the parasite species

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Autor(es): dc.creatorGraça, Grazielle Cardoso da-
Autor(es): dc.creatorVolpini, Angela Cristina-
Autor(es): dc.creatorRomero, Gustavo Adolfo Sierra-
Autor(es): dc.creatorOliveira Neto, Manoel Paes de-
Autor(es): dc.creatorHueb, Marcia-
Autor(es): dc.creatorPorrozzi, Renato-
Autor(es): dc.creatorBoité, Mariana Côrtes-
Autor(es): dc.creatorCupolillo, Elisa-
Data de aceite: dc.date.accessioned2021-10-14T18:24:56Z-
Data de disponibilização: dc.date.available2021-10-14T18:24:56Z-
Data de envio: dc.date.issued2017-12-07-
Data de envio: dc.date.issued2017-12-07-
Data de envio: dc.date.issued2012-08-
Fonte completa do material: dc.identifierhttp://repositorio.unb.br/handle/10482/28305-
Fonte completa do material: dc.identifierhttps://dx.doi.org/10.1590/S0074-02762012000500014-
Fonte: dc.identifier.urihttp://educapes.capes.gov.br/handle/capes/631089-
Descrição: dc.descriptionIn this study, PCR assays targeting different Leishmania heat-shock protein 70 gene (hsp70) regions, producing fragments ranging in size from 230-390 bp were developed and evaluated to determine their potential as a tool for the specific molecular diagnosis of cutaneous leishmaniasis (CL). A total of 70 Leishmania strains were analysed, including seven reference strains (RS) and 63 previously typed strains. Analysis of the RS indicated a specific region of 234 bp in the hsp70 gene as a valid target that was highly sensitive for detection of Leishmania species DNA with capacity of distinguishing all analyzed species, after polymerase chain reaction-restriction fragment length polymorfism (PCR-RFLP). This PCR assay was compared with other PCR targets used for the molecular diagnosis of leishmaniasis: hsp70 (1400-bp region), internal transcribed spacer (ITS)1 and glucose-6-phosphate dehydrogenase (G6pd). A good agreement among the methods was observed concerning the Leishmania species identification. Moreover, to evaluate the potential for molecular diagnosis, we compared the PCR targets hsp70-234 bp, ITS1, G6pd and mkDNA using a panel of 99 DNA samples from tissue fragments collected from patients with confirmed CL. Both PCR-hsp70-234 bp and PCR-ITS1 detected Leishmania DNA in more than 70% of the samples. However, using hsp70-234 bp PCR-RFLP, identification of all of the Leishmania species associated with CL in Brazil can be achieved employing a simpler and cheaper electrophoresis protocol.-
Formato: dc.formatapplication/pdf-
Idioma: dc.languageen-
Publicador: dc.publisherInstituto Oswaldo Cruz, Ministério da Saúde-
Direitos: dc.rightsAcesso Aberto-
Direitos: dc.rightsMemórias do Instituto Oswaldo Cruz - All the contents of this journal, except where otherwise noted, is licensed under a Creative Commons Attribution License (CC BY NC 4.0). Fonte: https://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762012000500014&lng=en&tlng=en. Acesso em: 04 dez. 2020.-
Palavras-chave: dc.subjectLeishmaniose-
Palavras-chave: dc.subjectDiagnóstico molecular-
Palavras-chave: dc.subjectIdentificação de espécies-
Palavras-chave: dc.subjectReação em cadeia de polimerase-
Título: dc.titleDevelopment and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identificatio nof the parasite species-
Tipo de arquivo: dc.typelivro digital-
Aparece nas coleções:Repositório Institucional – UNB

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