| Literature DB >> 1280640 |
T Urabe1, K Sano, M Tanno, J Mizoguchi, M Otani, M H Lee, T Takasaki, H Kusakabe, D T Imagawa, M Nakai.
Abstract
We developed a non-radioisotopic (non-RI) reverse transcriptase assay (RTA). The reverse transcriptase (RT) incorporates biotin-11-deoxyuridine-triphosphate (bio-dUTP) using a poly(rA) template hybridized with oligo(dT) primer that is immobilized on the surface of a 96-well microtiter plate. This assay is thus semi-automated by adapting it to an ELISA testing format. The incorporation of bio-dUTP was enhanced by adding cold dTTP to the reaction mixture, optimally in a molar ratio 4:1 (dTTP:bio-dUTP). This non-RI RTA is more sensitive than the conventional RI assay for the detection of purified Rous-associated virus 2 (RAV-2) and of human immunodeficiency virus type 1 (HIV-1) lysate. Because of its simple procedure, higher sensitivity and non-use of RI materials, the assay can be utilized not only for virological studies but also for routine safety screening of biological products for retroviral contamination.Entities:
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Year: 1992 PMID： 1280640 DOI： 10.1016/0166-0934(92)90063-j
Source DB: PubMed Journal: J Virol Methods ISSN： 0166-0934 Impact factor: 2.014