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Rota Virus

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Viruses come in different genomes and different forms however they all have a capsid where they maintain their genetic material. Viruses are classified into families that are distinguish based on their shape and form as well as they have names that are used for identification and is usually related to the type of disease they cause. Rotavirus is a member of the family Reoviridae that is nonenveloped segmented double stranded RNA and globally is the leading cause of severe diarrhea problems among infants and young children worldwide (1). Rotavirus developed in a wheel shaped virus that is surrounded by a triple layer coat composed of different structural proteins. The outer layer is composed of VP7 and VP4 that is cleaved by trypsin into two …show more content…

They used a rhesus monkey epithelial cell line MA104 and the murine fibroblast L929 cell line (ATCC) that were grown in advanced Eagle's minimal essential medium (MEM; Invitrogen) supplemented with 5% and 10% fetal bovine serum (FBS), respectively, at 37°C in a 5% CO2 atmosphere. Then, they propagated in the MA105 cells different kinds of RV strains. RV cell lysates were activated with trypsin. TLPs and DLPs of the RRV and BRV UK strains were purified by CsCl isopycnic gradients (45). The mammalian reovirus strain type 3 Dearing (ReoT3D) was propagated in L929 cells, and the viral titer was determined in MA104 cells (36, 46). Monoclonal antibodies were used for detection of cellular proteins. Virus infectivity was determined by either immunoperoxidase or immunofluorescence focus-forming assays. In order to bind the assays created they used the MA104 cells that where transferred with the indicated siRNAs. Confluent cells were then washed twice and incubated with MEM for 30 min at 37°C and virus binding was assayed. To evaluate the binding in the cells treated with neuraminidase, the assay was carried out. There were two type of treatments depending if the MA104 cells were LBPA-blocking assays they will incubate the cells of anti-LBPA antibody with an isotype IgG control or if the MA104 cells were protease inhibitors they were treated with leupeptin (in water) or with cathepsin B or L inhibitors. Then the cells that had grown on glass coverslips were washed twice with MEM to get them ready to localize the rotavirus particles with endocytic markers. The cells went through an immunoblot analysis. Finally the qRT-PCR was done as well as the statistical analysis using the two tail paired

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