AEGiS-11IAC: Endogenous HIV reverse transcriptase assays are more sensitive than other methods in determinations of HIV drug resistance.

11th International AIDS Conference

Vancouver, British Columbia — July 7-12, 1996

Endogenous HIV reverse transcriptase assays are more sensitive than other methods in determinations of HIV drug resistance.

Int Conf AIDS 1996 Jul 7-12; 11:438 (abstract no. Pub.A.1003)
Quan Y, Gu Z, Wainberg MA; McGill University AIDS Centre-Jewish General Hospital, Montreal, Canada. Fax: (514) 340-7537.

OBJECTIVE: To determine the utility of endogenous HIV reverse transcriptase (RT) assays in assays of drug resistance.

METHODS: Fully endogenous RT asays were performed using either wild-type (wt) or drug-resistant mutated viruses. These studies utilize the viral genome as template and are performed with lysed virions in the presence of all four nucleoside triphosphates (dNTP) and, as appropriate, nucleoside triphosphate inhibitors (ddNTP) of RT activity.

RESULTS: Viruses containing the M184V substitution were highly resistant to 3TC triphosphate, with an increase in IC50 of 250-fold, in comparison with wt recombinant virus. In regard to ddC triphosphate, the increase in IC50 conferred by either K65R or M184V recombinant virus was about 30-fold. These results are consistent with biochemical and tissue culture data. Recombinant viruses, containing both the K65R and M184 substitutions, displayed levels of resistance to each of 3TC-TP and ddC-TP at least 3-fold higher than those reported above.

CONCLUSIONS: Fully endogenous RT assays, as monitored by gel electrophoresis, may represent a more sensitive means of detecting drug resistance than either tissue culture or other cell-free biochemical methods.

Keywords: AEGIS, RNA-Directed DNA Polymerase, HIV, Lamivudine, Reverse Transcriptase Inhibitors, Drug Resistance, Anti-HIV Agents, Templates, Genetic, HIV-1, Zalcitabine, Pharmaceutical Preparations, DNA Primers, lamivudine triphosphate, methods, analysis, genetics, ICA11

960707
PubA1003