Author | Fumian, Tulio M. | |
Author | Leite, José Paulo G. | |
Author | Castello, Alejandro A. | |
Author | Gaggero, Aldo | |
Author | Caillou, Maria Susana L. de | |
Author | Miagostovich, Marize P. | |
Access date | 2018-03-20T14:51:33Z | |
Available date | 2018-03-20T14:51:33Z | |
Document date | 2010 | |
Citation | FUMIAN, Tulio M. et al. Detection of rotavirus A in sewage samples using multiplex qPCR and an evaluation of the ultracentrifugation and adsorption-elution methods for virus concentration. Journal of Virological Methods, v. 170, p. 42–46, 2010. | pt_BR |
ISSN | 0166-0934 | pt_BR |
URI | https://www.arca.fiocruz.br/handle/icict/25422 | |
Language | eng | pt_BR |
Publisher | Elsevier | pt_BR |
Rights | open access | pt_BR |
Subject in Portuguese | Rotavirus A | pt_BR |
Subject in Portuguese | Levivirus | pt_BR |
Subject in Portuguese | Ultracentrifugação | pt_BR |
Subject in Portuguese | Método de adsorção-eluição | pt_BR |
Subject in Portuguese | Reação em Cadeia da Polimerase Multiplex | pt_BR |
Title | Detection of rotavirus A in sewage samples using multiplex qPCR and an evaluation of the ultracentrifugation and adsorption-elution methods for virus concentration | pt_BR |
Type | Article | pt_BR |
DOI | 10.1016/j.jviromet.2010.08.017 | |
Abstract | Group A rotaviruses (RV-A) are the most common agents of viral gastroenteritis in children worldwide. The goal of this study was to compare two different methods to concentrate RV-A from sewage samples and to improve the detection and quantification of RV-A using a multiplex quantitative PCR assay with an internal control. Both RV-A and the internal control virus, bacteriophage PP7, were seeded into wastewater and then concentrated using either an ultrafiltration-based adsorption-elution protocol or an ultracentrifugation-based protocol. Real time multiplex quantitative PCR was used to quantify the purified RV-A and PP7, and the results of the multiplex assay were compared with the results of the monoplex assays. The ultracentrifugation-based method had a mean recovery rate of 47% (range: 34-60%), while the ultrafiltration-based adsorption-elution method had a mean recovery rate of 3.5% (range: 1.5-5.5%). These results demonstrate that ultracentrifugation is a more appropriate method for recovering RV-A from wastewater. This method together with the multiplex qPCR assay may be suitable for routine laboratory use. | 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 | Universidad Nacional de Quilmes Bernal. Laboratorio de Inmunología y Virología. Argentina. | pt_BR |
Affilliation | Universidad de Chile. Facultad de Medicina. ICBM. Programa de Virologia. Chile. | pt_BR |
Affilliation | Universidad de Tucumán. Instituto de Microbiología. Argentina. | 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 | PP7 | pt_BR |
Subject | Multiplex qPCR | pt_BR |
Subject | Ultracentrifugation | pt_BR |
Subject | Adsorption-elution method | pt_BR |
e-ISSN | 1879-0984 | |
Embargo date | 2013-01-01 | |