| Full text | |
| Author(s): |
Total Authors: 2
|
| Affiliation: | [1] State Univ Campinas UNICAMP, Inst Biol, Dept Genet Evolut & Bioagents, BR-13083970 Campinas, SP - Brazil
Total Affiliations: 1
|
| Document type: | Journal article |
| Source: | Journal of Virological Methods; v. 234, p. 156-159, AUG 2016. |
| Web of Science Citations: | 0 |
| Abstract | |
Rapid, sensitive and specific methods are necessary to detect and quantify infectious viruses. Cultivating and detecting enteric viruses in cell culture are difficult, thus impairing the advancement of knowledge regarding virus -induced diarrhea. Rotavirus (RV) detection has been conducted by serological or molecular biology methods, which do not provide information regarding viral infectivity. Molecular beacons (MBs) have demonstrated efficacy for viral detection in cell culture. We propose a MB assay to detect human rotavirus group A (HuRVA) in cell culture. MA104 cells were mock -infected or infected with HuRVA strains (RotaTeq vaccine and KS strains), and a specific MB for the HuRVA VP6 gene was used for virus detection. Mock -infected cells showed basal fluorescence, while infected cells exhibited increased fluorescence emission. MB hybridization to the viral mRNA target of HuRVA was confirmed. Fluorescence increased according to the increase in the number of infectious viral particles per cell (MOI 0.5-MOI 1). This technique provides quick and efficient HuRVA detection in cell culture without a need for viral culture for several days or many times until cytopathic effects are visualized. This methodology could be applied in the selection of samples for developing RV vaccines. (C) 2016 Elsevier B.V. All rights reserved. (AU) | |
| FAPESP's process: | 11/23865-1 - STUDY OF THE ACTION OF ROTAVIRUS NSP4 ON CELLULAR PERMEABILITY IN VITRO AND INTESTINAL FLUID ACCUMULATION IN RATS (MODEL IN VIVO) |
| Grantee: | Jéssica Wildgrube Bertol |
| Support Opportunities: | Scholarships in Brazil - Doctorate |