Abstract: Hantaviruses are members of the family Bunyaviridaeandare an emergingcause of diseaseworldwidewithhighlethalityin the Americas. In Brazil, thediagnosis forhantaviruses is basedonimmunologictechniquesassociatedwithconventionalRT-PCR.Anovelone-stepSYBRGreenreal-timeRT-PCRwasdevelopedfor the detectionandquantitationofAraraquarahantavirus(ARAV)and Rio Mamore hantavirus (RIOMV). Thedetectionlimitof assay was 10copies/µLofRNA in vitro transcribed of segment S. The specificity of assay was evaluatedbymeltingcurveanalysis, which showed thattheAraraquaravirusamplifiedproductgenerate dameltpeak at80.83±0.89°C without generating primer-dimersornon-specificproducts.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 ARAVand RIOMVand possibly other Brazilian hantaviruses.
Keywords: Hantaviruses; Araraquara virus; Rio Mamore virus; real-time quantitative RT-PCR; SYBR green I
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Machado, A.M.; de Souza, W.M.; de Pádua, M.; da Silva Rodrigues Machado, A.R.; Figueiredo, L.T.M. Development of a One-Step SYBR Green I Real-Time RT-PCR Assay for the Detection and Quantitation of Araraquara and Rio Mamore Hantavirus. Viruses 2013, 5, 2272-2281.
Machado AM, de Souza WM, de Pádua M, da Silva Rodrigues Machado AR, Figueiredo LTM. Development of a One-Step SYBR Green I Real-Time RT-PCR Assay for the Detection and Quantitation of Araraquara and Rio Mamore Hantavirus. Viruses. 2013; 5(9):2272-2281.
Machado, Alex M.; de Souza, William M.; de Pádua, Michelly; da Silva Rodrigues Machado, Aline R.; Figueiredo, Luiz T.M. 2013. "Development of a One-Step SYBR Green I Real-Time RT-PCR Assay for the Detection and Quantitation of Araraquara and Rio Mamore Hantavirus." Viruses 5, no. 9: 2272-2281.