| Journal: | Memorias do Instituto Oswaldo Cruz |
| Database: | PERIÓDICA |
| System number: | 000405572 |
| ISSN: | 0074-0276 |
| Authors: | Naveca, Felipe Gomes1 Nascimento, Valdinete Alves do1 Souza, Victor Costa de1 Nunes, Bruno Tardelli Diniz2 Rodrigues, Daniela Sueli Guerreiro2 Vasconcelos, Pedro Fernando da Costa2 |
| Institutions: | 1Fundacao Oswaldo Cruz, Instituto Leonidas e Maria Deane, Manaus, Amazonas. Brasil 2Secretaria de Vigilancia em Saude, Instituto Evandro Chagas, Ananindeua, Para. Brasil |
| Year: | 2017 |
| Season: | Jul |
| Volumen: | 112 |
| Number: | 7 |
| Pages: | 510-513 |
| Country: | Brasil |
| Language: | Inglés |
| Document type: | Artículo |
| Approach: | Experimental, aplicado |
| English abstract | We describe a sensitive method for simultaneous detection of Oropouche and Oropouche-like viruses carrying the Oropouche S segment, as well as the Mayaro virus, using a multiplexed one-step reverse transcription real-time polymerase chain reaction (RT-qPCR). A chimeric plasmid containing both Mayaro and Oropouche targets was designed and evaluated for the in vitro production of transcribed RNA, which could be easily used as a non-infectious external control. To track false-negative results due to PCR inhibition or equipment malfunction, the MS2 bacteriophage was also included in the multiplex assay as an internal positive control. The specificity of the multiplex assay was evaluated by Primer-Blast analysis against the entire GenBank database, and further against a panel of 17 RNA arboviruses. The results indicated an accurate and highly sensitive assay with amplification efficiency greater than 98% for both targets, and a limit of detection between two and 20 copies per reaction. We believe that the assay described here will provide a tool for Mayaro and Oropouche virus detection, especially in areas where differential diagnosis of Dengue, Zika and Chikungunya viruses should be performed |
| Disciplines: | Medicina |
| Keyword: | Diagnóstico, Virus, Técnicas de diagnóstico, Detección de virus, Reacción en cadena de la polimerasa (PCR), Virus Mayaro, Virus oropouche |
| Keyword: | Medicine, Diagnosis, Virus, Diagnostic techniques, Virus detection, Polymerase chain reaction (PCR), Mayaro virus, Oropouche virus |
| Full text: | Texto completo (Ver HTML) |
