Development of a One-Step SYBR Green I Real-Time RT-PCR Assay for the Detection and Quantitation of Araraquara and Rio Mamore Hantavirus

Autor(es)

Machado, Alex Martins ^[1] ; de Souza, William Marciel ^[1] ; de Padua, Michelly ^[1] ; da Silva Rodrigues Machado, Aline Rafaela ^[1] ; Moraes Figueiredo, Luiz Tadeu ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Sch Med Ribeirao Preto, Virol Res Ctr, BR-14049900 Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Viruses-Basel; v. 5, n. 9, p. 2272-2281, SEP 2013.
• Citações Web of Science: 12

Resumo

Hantaviruses are members of the family Bunyaviridae and are an emerging cause of disease worldwide with high lethality in the Americas. In Brazil, the diagnosis for hantaviruses is based on immunologic techniques associated with conventional RT-PCR. A novel one-step SYBR Green real-time RT-PCR was developed for the detection and quantitation of Araraquara hantavirus (ARAV) and Rio Mamore hantavirus (RIOMV). The detection limit of assay was 10copies/mu L of RNA in vitro transcribed of segment S. The specificity of assay was evaluated by melting curve analysis, which showed that the Araraquara virus amplified product generated a melt peak at 80.83 +/- 0.89 degrees C without generating primer-dimers or non-specific products. The assay was more sensitive than conventional RT-PCR and we detected two samples undetected by conventional RT-PCR. The one-step SYBR Green real-time quantitative RT-PCR is specific, sensible and reproducible, which makes it a powerful tool in both diagnostic applications and general research of ARAV and RIOMV and possibly other Brazilian hantaviruses. (AU)

• Processo FAPESP: 08/50617-6 - Estudos sobre vírus emergentes incluindo arbovirus, robovirus, vírus respiratórios e de transmissão congênita, no Centro de Pesquisa em Virologia da Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo
• Beneficiário: Luiz Tadeu Moraes Figueiredo
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 11/16866-1 - Pesquisa de anticorpos neutralizantes em pacientes com síndrome pulmonar e cardiovascular e roedores infectados por hantavírus
• Beneficiário: Michelly de Pádua
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 11/05067-0 - Estudo dos mecanismos de infecção e transmissão de hantavírus Araraquara em roedor-reservatório (Necromys lasiurus)
• Beneficiário: William Marciel de Souza
• Modalidade de apoio: Bolsas no Brasil - Mestrado