Psoralen monoadducted crosslinkable DNA oligonucleotide probes (COP's) to conserved regions of the HIV-I genome will be prepared, attached to polystyrene beads, and characterized for use in an innovative solution hybridization assay. The assay will be conducted at the melting temperature of the respective probe-target hybrid in the presence of high probe concentrations. Removal of excess probe and cellular debris will be achieved with stringent, denaturing washes. The capture, wash, and detection steps with the COP beads will be refined for use with multiwell microtiter plate technology. Clinical assay protocols employing the crosslinkable capture beads and a nonradioactive detection technique will be developed for the identification of latent, integrated or episomal HIV DNA in peripheral blood mononuclear cells. The polymerase chain reaction technique will be used to amplify a segment of HIV DNA for capture by the COP beads. Clinical assays employing the COP beads for the quantitative detection of HIV RNA will also be developed and characterized for use with patients undergoing therapeutic treatments for AIDS. An innovative, cascading nonradioactive amplification protocol for the RNA assay will be developed.
