Poly A-linked colorimetric microtiter plate assay for HIV reverse transcriptase.

K Suzuki; B P Craddock; N Okamoto; T Kano; R T Steigbigel

doi:10.1016/0166-0934(93)90054-u

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Poly A-linked colorimetric microtiter plate assay for HIV reverse transcriptase.

K Suzuki·B P Craddock·N Okamoto·T Kano·R T Steigbigel

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Abstract

An assay for detection of the reverse transcriptase (RT) of the human immunodeficiency virus (HIV) was developed using poly A linked to microtiter plate with colorimetric detection of incorporated biotin deoxyuridine triphosphate (biotin-dUTP). During the RT reaction, biotin-dUTP was incorporated into oligodeoxythymidylic acid (oligo-dT) which had been hybridized with poly A. At the detection step, horseradish peroxidase conjugated streptavidin was added, followed by the reaction of a colorimetric substrate for this enzyme. This method was contrasted with the two standard isotopic RT assays. There was excellent correlation between the colorimetric RT assay and each of two isotopic RT assays for both detection and quantification of avian myoblastosis virus reverse transcriptase (AMV-RT) and of HIV RT in human lymphocytes infected in vitro with HIV-1. The total assay required for performing the colorimetric assay, including the RT reaction, was 40 min.

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Biotin Colorimetry Deoxyuracil Nucleotides Digoxigenin Enzyme-Linked Immunosorbent Assay HIV Reverse Transcriptase HIV-1 Humans Indicators and Reagents Leukocytes, Mononuclear Poly A RNA-Directed DNA Polymerase Retroviridae Sensitivity and Specificity Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

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K Suzuki, B P Craddock, N Okamoto, T Kano, R T Steigbigel. 1993. Poly A-linked colorimetric microtiter plate assay for HIV reverse transcriptase.. https://doi.org/10.1016/0166-0934(93)90054-u

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