Sensitive phenotypic detection of minor drug-resistant human immunodeficiency virus type 1 reverse transcriptase variants.
Detection of drug-resistant variants is important for the clinical management of human immunodeficiency virus type 1 (HIV-1) infection and for studies on the evolution of drug resistance. Here we show that hybrid elements composed of the Saccharomyces cerevisiae retrotransposon Ty1 and the reverse transcriptase (RT) of HIV-1 are useful tools for detecting, monitoring, and isolating drug-resistant reverse transcriptases. This sensitive phenotypic assay is able to detect nonnucleoside reverse transcriptase inhibitor-resistant RT domains derived from mixtures of infectious molecular clones of HIV-1 in plasma and from clinical samples when the variants comprise as little as 0.3 to 1% of the virus population. Our assay can characterize the activities and drug susceptibilities of both known and novel reverse transcriptase variants and should prove useful in studies of the evolution and clinical significance of minor drug-resistant viral variants.
Nissley
Dwight V
DV
Basic Research Program, SAIC-Frederick, NCI Frederick, P.O. Box B, Frederick, MD 21702, USA. nissley@ncifcrf.gov
Halvas
Elias K
EK
Hoppman
Nicole L
NL
Garfinkel
David J
DJ
Mellors
John W
JW
Strathern
Jeffrey N
JN
D024882:Drug Resistance, Viral; D015658:HIV Infections / Q000188:drug therapy / Q000821:virology*; D054303:HIV Reverse Transcriptase / Q000187:drug effects* / Q000235:genetics; D015497:HIV-1 / Q000187:drug effects* / Q000201:enzymology* / Q000302:isolation & purification; D006801:Humans; D018626:Retroelements / Q000235:genetics; D018894:Reverse Transcriptase Inhibitors / Q000008:administration & dosage / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000235:genetics; D012680:Sensitivity and Specificity; D014170:Transformation, Genetic
D018626:Retroelements; D018894:Reverse Transcriptase Inhibitors; D054303:HIV Reverse Transcriptase