| Author | Fumian, Tulio M. | |
| Author | Leite, José Paulo G. | |
| Author | Rocha, Mônica S. | |
| Author | Andrade, Juliana S. R. | |
| Author | Fioretti, Julia M. | |
| Author | Assis, Rosane M. S. de | |
| Author | Assis, Matheus R. S. | |
| Author | Fialho, Alexandre M. | |
| Author | Miagostovich, Marise P. | |
| Access date | 2016-04-28T15:26:07Z | |
| Available date | 2016-04-28T15:26:07Z | |
| Document date | 2016 | |
| Citation | FUMIAN, Tulio M. et al. Performance of a one-step quantitative duplex RT-PCR for detection of rotavirus A and noroviruses GII during two periods of high viral circulation. Journal of Virological Methods, v.228, p.123-129, Fe. 2016. | pt_BR |
| ISSN | 0166-0934 | |
| URI | https://www.arca.fiocruz.br/handle/icict/14013 | |
| Language | eng | pt_BR |
| Publisher | Elsevier | pt_BR |
| Rights | restricted access | |
| Title | Performance of a one-step quantitative duplex RT-PCR for detection of rotavirus A and noroviruses GII during two periods of high viral circulation | pt_BR |
| Type | Article | |
| DOI | 10.1016/j.jviromet.2015.11.008 | |
| Abstract | Rotavirus A (RVA) and noroviruses (NoV) are the major viral agents of acute gastroenteritis (AGE) worldwide.
In the present study, we aimed to evaluate the performance of a one-step duplex quantitative
RT-PCR (dRT-qPCR) assay, established for detection and quantification of RVA and NoV genogroup II (GII)
using a single DNA standard curve (SC), as well as to investigate the association between fecal viral load
and optical density (OD) values, and viruses’ genotyping. The results obtained by dRT-qPCR in 530 fecal
samples from AGE cases were compared with methods employed for the diagnosis of those viruses as
follows: enzyme immunoassay (EIA) and polyacrylamide gel electrophoresis (PAGE) for RVA; and qualitative
PCR for NoV. By using dRT-qPCR, we detected RVA and NoV in 353 (66%), increasing the positivity
rate by 22.5% for RVA and 11.5% NoV, comparing the number of positive samples. RVA and NoV GII
were detected in a range of 5.17 × 103 to 6.56 × 109 and 3.76 × 103 to 9.13 × 1010 genome copies per
gram of feces, respectively. We observed a significant direct correlation between genome copies values
and optical density, using dRT-qPCR and EIA assays, respectively (Spearman = 0.41; p < 0.0001). Viruses
characterization demonstrated a predominance of NoV GII.4 Sidney 2012 variant during October 2013
to February 2014, followed by the emergence of RVA genotype G12P[8] in 2014. The established assay
using a single SC provides an early feedback concerning detection and quantification, with the advantage
of detecting simultaneously RVA and NoV GII, reducing time and reagent costs. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Subject | Rotavirus A | pt_BR |
| Subject | Noroviruses GII | pt_BR |
| Subject | Duplex quantitative RT-PCR | pt_BR |
| Subject | Genotyping | pt_BR |
| DeCS | Técnicas de Genotipagem | pt_BR |
| DeCS | Reação em Cadeia da Polimerase | pt_BR |
| DeCS | Rotavirus | pt_BR |
| DeCS | Norovirus | pt_BR |
| e-ISSN | 1879-0984 | |
