| Author | Filgueira, Camila Patrício | |
| Author | Moreira, Otacilio Cruz | |
| Author | Cantanhêde, Lilan Motta | |
| Author | Farias, Heloísa Martins Teixeira de | |
| Author | Porrozzi, Renato | |
| Author | Britto, Constança | |
| Author | Boité, Mariana Côrtes | |
| Author | Cupolillo, Elisa | |
| Access date | 2020-11-30T14:37:51Z | |
| Available date | 2020-11-30T14:37:51Z | |
| Document date | 2020 | |
| Citation | FILGUEIRA, Camilia Patrício Braga et al. Comparison and clinical validation of qPCR assays targeting Leishmania 18S rDNA and HSP70 genes in patients with American Tegumentary Leishmaniasis. PLoS Negl Trop Dis., v. 14, n. 10., e0008750, 18p, Oct. 2020. | pt_BR |
| ISSN | 1935-2727 | pt_BR |
| URI | https://www.arca.fiocruz.br/handle/icict/44675 | |
| Language | eng | pt_BR |
| Publisher | Public Lybrary of Science | pt_BR |
| Rights | open access | |
| Subject in Portuguese | PCR | pt_BR |
| Subject in Portuguese | Comparação e validação clínica | pt_BR |
| Subject in Portuguese | rDNA de Leishmania 18S | pt_BR |
| Subject in Portuguese | Genes HSP70 | pt_BR |
| Subject in Portuguese | Pacientes | pt_BR |
| Subject in Portuguese | Leishmaniose tegumentar americana | pt_BR |
| Title | Comparison and clinical validation of qPCR assays targeting Leishmania 18S rDNA and HSP70 genes in patients with American Tegumentary Leishmaniasis | pt_BR |
| Type | Article | |
| DOI | 10.1371/journal. pntd.0008750 | |
| Abstract | Leishmaniasis is a worldwide neglected disease, encompassing asymptomatic infections
and different clinical forms, such as American Tegumentary Leishmaniasis (ATL) which is
part of the complex of diseases caused by protozoan parasites from Leishmania genus,
transmitted by sand fly vectors. As a neglected disease, much effort is still needed in treatment
and diagnosis. Currently, ATL diagnosis is mainly made by parasite detection by
microscopy. The sensitivity of the method varies, and factors such as collection procedures
interfere. Molecular approaches, specially based on Real Time PCR (qPCR) technique, has
been widely used to detect Leishmania infection and to quantify parasite load, once it is a
simple, rapid and sensitive methodology, capable to detect low parasite concentrations and
less prone to variability. Although many studies have been already published addressing
the use of this technique, an improvement on these methodologies, including an analytical
validation, standardization and data association is demanded. Moreover, a proper validation
by the assay by the use of clinical samples is still required. In this sense, the purpose of the
present work is to compare the performance of qPCR using two commonly used targets
(18S rDNA and HSP70) with an internal control (RNAse P) in multiplex reactions. Additionally,
we validated reactions by assaying 88 samples from patients presenting different clinical
forms of leishmaniasis (cutaneous, mucosal, recent and old lesions), representing the
diversity found in Brazil’s Amazon Region. Following the methodology proposed herein, the
results indicate the use of both qPCR assays, 18S rDNA and HSP70, to achieve a very
good net sensitivity (98.5%) and specificity (100%), performing simultaneous or sequential
testing, respectively. With this approach, our main goal is to conclude the first step of a further
multicenter study to propose the standardization of detection and quantification of
Leishmania. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Epidemiologia Genética. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Subject | PCR | pt_BR |
| Subject | American Tegumentary Leishmaniasis | pt_BR |
| Subject | Leishmania 18S rDNA and HSP70 genes | pt_BR |
| Subject | qPCR assays | pt_BR |
| Subject | Comparison and clinical validation | pt_BR |
| Subject | Patients | pt_BR |
| e-ISSN | 1935-2735 | |
