ABSTRACT
We report conditions for the growth of human faecal astrovirus in a continuous colonic carcinoma cell line (CaCo-2). Purified particles contained three polypeptides, one of which (24k) appeared loosely held on the exterior.
Subject(s)
Feces/microbiology , Mamastrovirus/growth & development , Virus Cultivation , Viruses, Unclassified/growth & development , Animals , Cytopathogenic Effect, Viral , Diarrhea/microbiology , Electrophoresis, Polyacrylamide Gel , HeLa Cells , Humans , Mamastrovirus/ultrastructure , Tumor Cells, Cultured , Vero Cells , Viral Proteins/analysisABSTRACT
Adenovirus DNA was isolated directly from virus-containing stools and digested with restriction endonucleases. The resulting fragments were separated by polyacrylamide gel electrophoresis (PAGE) and visualized by silver staining. This enabled us to assign most of the viruses detected to subgenus, serotype and, sometimes, unique strains. Although less sensitive than electron microscopy, the method allowed more information about the infecting virus to be obtained and no cultivation was necessary. Comparison with culture also allowed dual infections to be recognized. A 2-year survey of faecal adenoviruses in Newcastle upon Tyne showed that type 41 (strain 41a) was the predominant type and strain 41p was not recorded. Heterogeneity in strain 41a was also noted as found elsewhere. Adenovirus type 40 was common prior to 1985 but was absent during the last 2 years.
Subject(s)
Adenoviruses, Human/classification , DNA, Viral/analysis , Feces/microbiology , Adenoviruses, Human/genetics , DNA Restriction Enzymes , Deoxyribonucleases, Type II Site-Specific , Electrophoresis, Polyacrylamide Gel , Humans , Predictive Value of Tests , Silver , Staining and Labeling , United KingdomABSTRACT
A commercially available latex agglutination test, Rotalex (Orion Diagnostics, Finland), for detecting rotaviruses was evaluated in comparison with four other tests (electron microscopy, immunofluorescence, polyacrylamide gel electrophoresis, and enzyme linked immunosorbent assay) routinely used in our laboratories. Although Rotalex was the least complex method, it showed lack of specificity and sensitivity when carried out according to the manufacturer's instructions. Four basic modifications of Rotalex are described. These include the use of Hank's balanced salt solution, increasing the incubation time to 20 min, reading the agglutination result by an experienced observer, and the use of 50 mm square glass plates. The modified procedure gave results which were comparable with those obtained by electron microscopy, immunofluorescence, polyacrylamide gel electrophoresis, and enzyme linked immunosorbent assay. The latter techniques, when used to detect rotavirus, all gave similar results.
