This application will determine the feasibility of a new phenotypic assay for drug susceptibility of HIV-1 protease. The technique will combine two procedures, one developed at Sepracor to assess HIV protease drug resistance, and one developed at the CDC for detection of extremely minute quantities of reverse transcriptase (Amp-RT assay). Virioin- derived HIV protease from patient samples will act on a precursor polypeptide to release active reverse transcriptase. The released reverse transcriptase will then be quantified using Amp-RT. Inclusion of protease inhibitors in the assay will determine which inhibitors still have activity against virion-associated protease. The applicant contends that such a tool would be of immense value in the custom design of effective combination therapy for AIDS patients.
Thesaurus Terms:AIDS therapy, antiAIDS agent, diagnosis design /evaluation, diagnosis quality /standard, drug screening /evaluation, enzyme activity, human immunodeficiency virus 1, method development, protease inhibitor RNA directed DNA polymerase, phenotype, rapid diagnosis, virus protein, viruslike particle enzyme linked immunosorbent assay, genotype, polymerase chain reaction, southern blottingNational Institute of Allergy and Infectious Diseases (NIAID)