Please use this identifier to cite or link to this item:
https://www.arca.fiocruz.br/handle/icict/25422
Type
ArticleCopyright
Open access
Embargo date
2013-01-01
Collections
- IOC - Artigos de Periódicos [12973]
Metadata
Show full item record
DETECTION OF ROTAVIRUS A IN SEWAGE SAMPLES USING MULTIPLEX QPCR AND AN EVALUATION OF THE ULTRACENTRIFUGATION AND ADSORPTION-ELUTION METHODS FOR VIRUS CONCENTRATION
Levivirus
Ultracentrifugação
Método de adsorção-eluição
Reação em Cadeia da Polimerase Multiplex
Author
Affilliation
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro, RJ. Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro, RJ. Brasil.
Universidad Nacional de Quilmes Bernal. Laboratorio de Inmunología y Virología. Argentina.
Universidad de Chile. Facultad de Medicina. ICBM. Programa de Virologia. Chile.
Universidad de Tucumán. Instituto de Microbiología. Argentina.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro, RJ. Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro, RJ. Brasil.
Universidad Nacional de Quilmes Bernal. Laboratorio de Inmunología y Virología. Argentina.
Universidad de Chile. Facultad de Medicina. ICBM. Programa de Virologia. Chile.
Universidad de Tucumán. Instituto de Microbiología. Argentina.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro, RJ. Brasil.
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.
Keywords in Portuguese
Rotavirus ALevivirus
Ultracentrifugação
Método de adsorção-eluição
Reação em Cadeia da Polimerase Multiplex
Share