Rapid, simple method of preparing rotaviral double-stranded ribonucleic acid for analysis by polyacrylamide gel electrophoresis.
A procedure for extracting rotaviral double-stranded ribonucleic acid (RNA) directly from fecal and intestinal specimens collected from calves and pigs is described. This procedure provides a rapid, simple, reproducible method of obtaining rotaviral double-stranded RNA preparations suitable for electrophoretic analysis in polyacrylamide-agarose composite gels. The rotaviral genome electrophoretic migration pattern produced by double-stranded RNA extracted directly from a specimen by this procedure was qualitatively identical to the electrophoretic migration pattern obtained with double-stranded RNA extracted from purified rotavirus derived from the same specimen. Direct extraction of specimens containing porcine rotavirus-like virus by this procedure gave preparations that had electrophoretic migration patterns similar, but not identical, to the characteristic electrophoretic migration pattern of the rotaviral genome. Sufficient rotaviral double-stranded RNA could be extracted from 6 ml of fecal or intestinal specimen by this procedure to permit 15 or more electrophoretic assays.
D000818:Animals; D002417:Cattle / Q000382:microbiology; D004591:Electrophoresis, Polyacrylamide Gel; D005243:Feces / Q000382:microbiology; D007422:Intestines / Q000382:microbiology; D012330:RNA, Double-Stranded / Q000032:analysis / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification*; D012087:Reoviridae / Q000032:analysis*; D012401:Rotavirus / Q000032:analysis*; D013552:Swine / Q000382:microbiology
D012330:RNA, Double-Stranded; D012367:RNA, Viral