| Author | Graça, Grazielle Cardoso da | |
| Author | Volpini, Angela Cristina | |
| Author | Romero, Gustavo Adolfo Sierra | |
| Author | Oliveira Neto, Manoel Paes de | |
| Author | Hueb, Marcia | |
| Author | Porrozzi, Renato | |
| Author | Boité, Mariana Côrtes | |
| Author | Cupolillo, Elisa | |
| Access date | 2014-04-01T13:05:21Z | |
| Available date | 2014-04-01T13:05:21Z | |
| Document date | 2012 | |
| Citation | GRAÇA, Grazielle Cardoso da et al. Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species. Memórias do Instituto Oswaldo Cruz. Rio de Janeiro, v. 107, n. 5, p. 664-674, 2012. | pt_BR |
| ISSN | 0074-0276 | |
| URI | https://www.arca.fiocruz.br/handle/icict/7478 | |
| Sponsorship | FIOCRUZ, FAPERJ, CNPq (MCT/CNPq/CT-Saúde/MS/SCTIE/DECIT 34/2008 - Doenças Negligenciadas - 576104/2008-6), Edital Pró-Equipamentos CAPES (27/2010), European Union (LeishEpinetSA, INCO-CT2005-015407) GC da G and ACV contributed equally to this work. | pt_BR |
| Language | eng | pt_BR |
| Publisher | Fundação Oswaldo Cruz | pt_BR |
| Rights | open access | |
| Title | Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species | pt_BR |
| Type | Article | |
| DOI | 10.1590/S0074-02762012000500014 | |
| Abstract | In 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. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Coleção de Leishmania. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Coleção de Leishmania. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Grupo de Genômica e Biologia Computacional. Belo Horizonte, MG, Brasil. | pt_BR |
| Affilliation | Universidade de Brasília. Núcleo de Medicina Tropical. Laboratório de Leishmanioses Brasília, DF, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Universidade Federal de Mato Grosso. Faculdade de Medicina. Cuiabá, MT, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Coleção de Leishmania. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Coleção de Leishmania. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Coleção de Leishmania. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Subject | Leishmaniasis | pt_BR |
| Subject | Molecular diagnosis | pt_BR |
| Subject | Species identification | pt_BR |
| Subject | Polymerase chain reaction | pt_BR |
| Subject | RFLPs | pt_BR |
| Subject | hsp70 | pt_BR |
