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Updated: Aug 12, 2026

Rapid Screening of HIV Reverse Transcriptase and Integrase Inhibitors
Published on: April 10, 2014
A high-throughput homogeneous assay for reverse transcriptase using generic reagents and time-resolved fluorescence
J H Zhang1, T Chen, S H Nguyen
1DuPont Pharmaceuticals Research Laboratories, Wilmington, Delaware 19880-0400, USA. ji-hu.zhang@dupontpharma.com
Abstract:
A homogeneous time-resolved fluorescence (HTRF) assay has been developed for determining the activity of HIV reverse transcriptase (HIV-RT). By using a sequential capping strategy, the assay has been configured to utilize only generic reagents such as biotinylated dUTP, streptavidin-allophycocyanin, and streptavidin-europium. The assay was optimized for a HIV-RT high-throughput screen. Under optimized conditions, a signal-to-background ratio of approximately 10:1 and a Z' factor of 0.8 were obtained. The titration curves of several known HIV-RT inhibitors have been evaluated with this assay format. This HTRF format can be used for high-throughput assays with other nucleotide polymerases.

