Described is a process for detecting reverse transcriptase activity and, thereby, reverse transcriptase inhibitors using fluorescence polarization, comprising, mixing a DNA primer with an RNA template. Then forming an RNA/DNA duplex utilizing the reverse transcriptase and removing the RNA from the RNA/DNA duplex to form single-stranded DNA. Finally, adding a fluorescent-labeled oligonucleotide complementary to the single-stranded DNA for hybridizing to the single-stranded DNA; and, measuring the fluorescence polarization.