Multiplexed reverse transcription real-time polymerase chain reaction for simultaneous detection of Mayaro, Oropouche, and Oropouche-like viruses

dc.creatorNaveca, Felipe Gomes
dc.creatorNascimento, Valdinete Alves do
dc.creatorSouza, Victor Costa de
dc.creatorNunes, Bruno Tardelli Diniz
dc.creatorRodrigues, Daniela Sueli Guerreiro
dc.creatorVasconcelos, Pedro Fernando da Costa
dc.date2017-06-13T14:42:56Z
dc.date2017-06-13T14:42:56Z
dc.date2017
dc.date.accessioned2023-09-26T20:39:39Z
dc.date.available2023-09-26T20:39:39Z
dc.identifierNAVECA, Felipe Gomes; et al. Multiplexed reverse transcription real-time polymerase chain reaction for simultaneous detection of Mayaro, Oropouche, and Oropouche-like viruses. Mem. Inst. Oswaldo Cruz, Rio de Janeiro , v. 112, n. 7, p. 510-513, July 2017.
dc.identifier0074-0276
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/19371
dc.identifier10.1590/0074-02760160062
dc.identifier1678-8060
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8861514
dc.descriptionWe 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.
dc.formatapplication/pdf
dc.languagepor
dc.publisherInstituto Oswaldo Cruz, Ministério da Saúde
dc.rightsopen access
dc.subjectReal-time PCR
dc.subjectMayaro Virus
dc.subjectOropouche Virus
dc.subjectAmazon
dc.titleMultiplexed reverse transcription real-time polymerase chain reaction for simultaneous detection of Mayaro, Oropouche, and Oropouche-like viruses
dc.typeArticle


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