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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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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Author(s):
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]
Total Authors: 5
Affiliation:
[1] Univ Sao Paulo, Sch Med Ribeirao Preto, Virol Res Ctr, BR-14049900 Sao Paulo - Brazil
Total Affiliations: 1
Document type: Journal article
Source: Viruses-Basel; v. 5, n. 9, p. 2272-2281, SEP 2013.
Web of Science Citations: 12
Abstract

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)

FAPESP's process: 11/05067-0 - Study of the mechanisms of infection and transmission of hantaviruses in rodent reservoir Araraquara (Necromys lasiurus)
Grantee:William Marciel de Souza
Support type: Scholarships in Brazil - Master
FAPESP's process: 08/50617-6 - Studies on emerging viruses including arbovirus, robovirus, respiratory viruses and congenital transmission, at the Centro de Pesquisa em Virologia da Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo
Grantee:Luiz Tadeu Moraes Figueiredo
Support type: Research Projects - Thematic Grants
FAPESP's process: 11/16866-1 - Research of neutralizing antibodies in patients with Hantavirus pulmonary and cardiovascular syndrome and rodents infected with Araraquara Hantavirus
Grantee:Michelly de Pádua
Support type: Scholarships in Brazil - Master