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

Details Rapid single-cycle assay for human immunodeficiency virus type-1 Rapid single-cycle assay for human immunodeficiency virus type-1

1998-08-06

2000-08-15

Stucker, Jeffrey

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving virus or bacteriophage

435 6, 4352351, C12Q 170

Patent

active

061034622

ABSTRACT:
An in vitro, single cycle, recombinant virus assay (RVA) for determining inhibition of HIV replication by a protease inhibitor 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; 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.

REFERENCES:
Kellam et al. "Recombinant Virus Assay: a Rapid, Phenotypic Assay for Assessment of Drug Susceptibility of Human Immunodeficiency Virus Type 1 Isolates", Antimicrobial Agents and Chemotherapy, vol. 38, No. 1 (Jan. 1994), pp. 23-30.
Harrigan et al. "Significance of Amino Acid Variation at Human Immunodeficiency Virus Type 1 Reverse Transcriptase Residue 210 for Zidovudine Susceptibility", Journal of Virology, vol. 70, No. 9 (1996), pp. 5930-5934.

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