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

Figure 1

Schematic diagram showing the principle of the 3′-processing integrase activity assay based on real-time PCR. HIV-1 LTR substrate is labelled with biotin at 3′ end of sense strand. IN has activity to cleave dinucleotide from the LTR oligonucleotide at 3′ end at CA recognition motif. After washing step, only the processed biotinylated GT-dinucleotide (pink colour) binds to avidin-coated tube. The unprocessed biotinylated HIV-1 LTR substrates regarding IN activity can be amplified and detected by specific primers and probe. Early signal for unprocessed reaction is represented by blue line, whereas late signal for processed reaction is represented by red line.