Viruses 2013, 5(9), 2272-2281; doi:10.3390/v5092272
Article

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

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Received: 13 July 2013; in revised form: 10 September 2013 / Accepted: 17 September 2013 / Published: 19 September 2013
(This article belongs to the Special Issue Hantaviruses)
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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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MDPI and ACS Style

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.

AMA Style

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.

Chicago/Turabian Style

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.


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