A Novel Phenotypic Drug Susceptibility Assay for Human Immunodeficiency Virus Type 1

doi:10.1128/AAC.44.4.920-928.2000

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Antimicrobial Agents and Chemotherapy, April 2000, p. 920-928, Vol. 44, No. 4
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

A Novel Phenotypic Drug Susceptibility Assay for Human Immunodeficiency Virus Type 1

Christos J. Petropoulos,^* Neil T. Parkin, Kay L. Limoli, Yolanda S. Lie, Terri Wrin, Wei Huang, Huan Tian, Douglas Smith, Genine A. Winslow, Daniel J. Capon, and Jeannette M. Whitcomb

ViroLogic, Inc., South San Francisco, California 94080

Received 27 September 1999/Returned for modification 1 December 1999/Accepted 5 January 2000

Although combination antiretroviral therapy has resulted in a considerable improvement in the treatment of human immunodeficiencyvirus (HIV) type 1 (HIV-1) infection, the emergence of resistantvirus is a significant obstacle to the effective management ofHIV infection and AIDS. We have developed a novel phenotypic drugsusceptibility assay that may be useful in guiding therapy andimproving long-term suppression of HIV replication. Susceptibilityto protease (PR) and reverse transcriptase (RT) inhibitors ismeasured by using resistance test vectors (RTVs) that containa luciferase indicator gene and PR and RT sequences derived fromHIV-1 in patient plasma. Cells are transfected with RTV DNA, resultingin the production of virus particles that are used to infect targetcells. Since RTVs are replication defective, luciferase activityis measured following a single round of replication. The assayhas been automated to increase throughput and is completed in8 to 10 days. Test results may be useful in facilitating the selectionof optimal treatment regimens for patients who have failed priortherapy or drug-naive patients infected with drug-resistant virus.In addition, the assay can be used to evaluate candidate drugsand assist in the development of new drugs that are active againstresistant strains of HIV-1.

^* Corresponding author. Mailing address: 270 East Grand Ave., South San Francisco, CA 94080. Phone: (650) 866-7439. Fax: (650)635-1111. E-mail: cpetropoulos{at}virologic.com.

Present address: Genetic Information Systems, Inc., Hillsborough, CA94010.

Antimicrobial Agents and Chemotherapy, April 2000, p. 920-928, Vol. 44, No. 4
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

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