Novel 3′-Processing Integrase Activity Assay by Real-Time PCR for Screening and Identification of HIV-1 Integrase Inhibitors

Figure 5

The 3′-processing IN activity assay by real-time PCR. Biotinylated HIV-1 LTR substrates were incubated with or without IN enzyme in a reaction tube at 37°C for 16 h. The reaction was transferred to an avidin-coated real-time PCR tube. After washing 3 times, the remaining of unprocessed biotinylated HIV-1 LTR substrates was then amplified and detected by specific primers and probe as described above. The reaction mixture without IN is shown in green line. The reaction mixture with IN is shown in blue line. Negative control is shown in black line. Ct: cycle threshold.