Rapid single-cycle assay for human immunodeficiency virus type 1 drug resistance

摘要

The invention provides an in vitro single cycle, recombinant virus assay (RVA) for determining inhibition of HIV replication by a protease inhibitor. The assay comprises transfecting a human epithelial cell line with amplified HIV protease sequences of an HIV virus; an HIV envelope defective molecular clone having a complete deletion of its protease coding sequence as well as two separate deletions in its env gene and a deletion of part of its gag gene; and a plasmid containing HIV envelope coding sequence under the control of a promoter for phenotypic complementation of the envelope defective molecular clone. The transfected cells are cultured in the presence of a protease inhibitor to produce a testable stock of infectious particles that can be used to infect indicator cells containing an indicator gene without amplification of the infectious particles prior to infecting the indicator cells. Accumulation of indicator gene product is a measure of inhibition of HIV replication by the protease inhibitor.