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pubmed-article:11424108pubmed:abstractTextConventional phenotypic analysis of resistance of the human immunodeficiency virus (HIV) to antiviral therapy is time-consuming and requires culture of infectious virus. Although phenotypic analyses may be desirable, rapid generation of test results and decentralized availability of the test system will be important to achieve utility in the clinical practice. This study describes the design of an alternative phenotypic resistance test using replication incompetent viral vectors. Chimeric HIV vectors containing a marker gene were generated. The env and most of the regulatory and accessory genes of HIV were removed. In addition, the 3'U3 region was deleted to obtain a self-inactivating construct. Cotransfection of the plasmid with a plasmid that provided the vesicular stomatitis virus glycoprotein resulted in the production of replication-incompetent virus vectors. Infection of susceptible cells with the vectors led to marker gene expression. Vector production in the presence of protease (PR) inhibitors, or infection in the presence of reverse transcriptase (RT) or integrase (IN) inhibitors reduced marker gene expression in a dose-dependent manner. Marker gene activity was preserved at higher drug levels if vectors contained RT and PR genes from resistant virus isolates. Sensitivity to nucleoside and non-nucleoside RT inhibitors, protease and integrase inhibitors could be determined in 10 working days. The phenotypic drug resistance test using replication-incompetent HIV vectors significantly speeds up drug resistance measurements and allows testing at reduced biosafety levels. This will make clinical use of phenotypic assessment of antiviral resistance more feasible.lld:pubmed
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pubmed-article:11424108pubmed:authorpubmed-author:RethwilmAAlld:pubmed
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pubmed-article:11424108pubmed:copyrightInfoCopyright 2001 Wiley-Liss, Inc.lld:pubmed
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pubmed-article:11424108pubmed:volume64lld:pubmed
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pubmed-article:11424108pubmed:pagination223-31lld:pubmed
pubmed-article:11424108pubmed:dateRevised2007-11-15lld:pubmed
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pubmed-article:11424108pubmed:year2001lld:pubmed
pubmed-article:11424108pubmed:articleTitlePhenotypic analysis of the sensitivity of HIV-1 to inhibitors of the reverse transcriptase, protease, and integrase using a self-inactivating virus vector system.lld:pubmed
pubmed-article:11424108pubmed:affiliationInstitute for Virology and Immunobiology, Julius Maximilians University, Würzburg, Germany.lld:pubmed
pubmed-article:11424108pubmed:publicationTypeJournal Articlelld:pubmed
pubmed-article:11424108pubmed:publicationTypeResearch Support, Non-U.S. Gov'tlld:pubmed
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