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| Autor | Albuquerque, Suênia da Cunha Gonçalves de | pt_BR |
| Autor | Silva, Rômulo Pessoa e | pt_BR |
| Autor | Morais, Rayana Carla Silva de | pt_BR |
| Autor | Silva, Lays Adrianne Mendonça Trajano | pt_BR |
| Autor | Silva, Carlos Gustavo Régis da | pt_BR |
| Autor | Brandão Filho, Sinval Pinto | pt_BR |
| Autor | Cavalcanti, Milena de Paiva | pt_BR |
| Fecha de acceso | 2017-11-24T13:40:36Z | |
| Fecha de disponibilización | 2017-11-24T13:40:36Z | |
| Fecha de publicación | 2014 | |
| Referencia | ALBUQUERQUE, Suênia da Cunha Gonçalves de et al. Tracking false-negative results in molecular diagnosis: proposal of a triplex-PCR based method for leishmaniasis diagnosis. Journal of Venomous Animals and Toxins including Tropical Diseases, v. 20, n. 16, p. 1-6, 22 Apr. 2014. | pt_BR |
| ISSN | 1678-9199 | pt_BR |
| URI | https://www.arca.fiocruz.br/handle/icict/23341 | |
| Descripción | Carlos Gustavo Régis da Silva - Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil. Documento produzido em parceria ou por autor vinculado à Fiocruz, mas não consta a informação no documento. | pt_BR |
| Idioma | eng | en_US |
| Editor | BMC | pt_BR |
| Derechos de autor | open access | pt_BR |
| Palabras clave en Portugués | Controle de extração | pt_BR |
| Palabras clave en Portugués | Reação em Cadeia da Polimerase Multiplex | pt_BR |
| Palabras clave en Portugués | pUC18 | pt_BR |
| Palabras clave en Portugués | Leishmaniose | pt_BR |
| Palabras clave en Portugués | Diagnóstico | pt_BR |
| Palabras clave en Portugués | Resultado falso negativo | pt_BR |
| Título | Tracking false-negative results in molecular diagnosis: proposal of a triplex-PCR based method for leishmaniasis diagnosis | en_US |
| Tipo del documento | Article | pt_BR |
| DOI | 10.1186/1678-9199-20-16 | pt_BR |
| Resumen en Inglés | Background: Molecular biological methods have become increasingly relevant to the diagnosis and control of infectious diseases, such as leishmaniasis. Since various factors may affect the sensitivity of PCR assays, including DNA yield and purity, an optimal extraction method is pivotal. Losses of a parasite’s DNA during extraction may significantly impair its detection by PCR and lead to false-negative results. This study proposes a triplex PCR assay targeting the parasite’s DNA, an external control (pUC18) and an internal control (G3PD) for accurate diagnosis of leishmaniasis. Results: Two primer pairs were designed to detect the plasmid pUC18 and a triplex PCR assay targeting the Leishmania braziliensis kinetoplast DNA, the external control and the internal control was standardized. The triplex PCR assay was assessed for its ability to detect the three target DNA fragments simultaneously. PCR products from pUC18 DNA resulted in bands of 368 (P1) and 316 (P2) base pairs (bp). The triplex PCR optimized with the chosen external control system (P1) allowed the simultaneous detection of the internal control (G3PD – 567 bp) as well as of small quantities (10 pg) of the target parasite’s DNA, detected by amplification of a 138 bp product. Conclusions: The new tool standardized herein enables a more reliable interpretation of PCR results, mainly by contributing to quality assurance of leishmaniasis diagnosis. Furthermore, after simple standardization steps, this protocol could be applied to the diagnosis of other infectious diseases in reference laboratories. This triplex PCR enables the assessment of small losses during the DNA extraction process, problems concerning DNA degradation (sample quality) and the detection of L. braziliensis kDNA. | en_US |
| Afiliación | Fundaçao Oswaldo Cruz. Instituto Aggeu Magalhães. Departamento de Imunologia. Recife, PE, Brasil. | pt_BR |
| Afiliación | Fundaçao Oswaldo Cruz. Instituto Aggeu Magalhães. Departamento de Imunologia. Recife, PE, Brasil. | pt_BR |
| Afiliación | Fundaçao Oswaldo Cruz. Instituto Aggeu Magalhães. Departamento de Imunologia. Recife, PE, Brasil. | pt_BR |
| Afiliación | Fundaçao Oswaldo Cruz. Instituto Aggeu Magalhães. Departamento de Imunologia. Recife, PE, Brasil. | pt_BR |
| Afiliación | Fundaçao Oswaldo Cruz. Instituto Aggeu Magalhães. Departamento de Imunologia. Recife, PE, Brasil. | pt_BR |
| Afiliación | Fundaçao Oswaldo Cruz. Instituto Aggeu Magalhães. Departamento de Imunologia. Recife, PE, Brasil. | pt_BR |
| Afiliación | Fundaçao Oswaldo Cruz. Instituto Aggeu Magalhães. Departamento de Imunologia. Recife, PE, Brasil. | pt_BR |
| Palavras clave en Inglês | Extraction control | en_US |
| Palavras clave en Inglês | Multiplex PCR | en_US |
| Palavras clave en Inglês | pUC18 | en_US |
| Palavras clave en Inglês | Leishmaniasis | en_US |
| Palavras clave en Inglês | Diagnosis | en_US |
| Palavras clave en Inglês | False-negative result | en_US |
| DeCS | Animais | pt_BR |
| DeCS | Diagnóstico | pt_BR |
| DeCS | Reações Falso-Negativas | pt_BR |
| DeCS | Reação em Cadeia da Polimerase | pt_BR |
| DeCS | Leishmaniose | pt_BR |
| DeCS | patologia | pt_BR |
| e-ISSN | 1678-9199 | pt_BR |
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