Rapid, simple method of preparing rotaviral double-stranded ribonucleic acid for analysis by polyacrylamide gel electrophoresis

Authors:
  • KTK W Theil
    K W Theil
  • CMC M McCloskey
    C M McCloskey
  • LSL J Saif
    L J Saif
  • DRD R Redman
    D R Redman
  • EBE H Bohl
    E H Bohl
  • DHD D Hancock
    D D Hancock
  • EKE M Kohler
    E M Kohler
  • PMP D Moorhead
    P D Moorhead
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Abstract
A procedure for extracting rotaviral double-stranded 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 speciment. 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.

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