title: A simple and rapid reverse transcriptase assay for the detection of retroviruses in cell cultures.
abstract: Reverse transcriptase (RT) is a good diagnostic tool for the detection of retroviruses. We have developed a simple and rapid assay for RT activity in culture supernatants. A 370-base RNA sequence from the tetracycline-resistance gene in pBR322 plasmid DNA was used as a template for RT-mediated cDNA synthesis. To detect the resultant cDNA, we used the nested polymerase chain reaction. A sensitivity test using purified recombinant RT of human immunodeficiency virus type 1 demonstrated that the detection limit of this method was 10-7-10-8 units of RT activity in 20 mul of a test sample (2 x 10-9-2 x 10-10 units ml-1). This method detected RT activity in unconcentrated supernatants of cell cultures infected with human T-cell leukemia virus, Moloney murine leukemia virus, Moloney murine sarcoma virus, or Rous sarcoma virus. This nonisotopic method provides results within 10 h and is useful for quality control to detect retroviruses in cell cultures.
authors.affiliation: Animal Cell Bank, Institute for Fermentation, Osaka (IFO), 2-17-85, Juso-honmachi, Yodogawa-ku, Osaka, 532, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Kuno
authors.forename: H
authors.initials: H
authors.lastname: Ikeda
authors.forename: M
authors.initials: M
authors.lastname: Takeuchi
authors.forename: T
authors.initials: T
authors.lastname: Yoshida