Mutations detection in HIV-1 protease gene through a sequencing method using digoxigenin labeled primers

Development of drug-resistant mutants is a general problem in antiretrovira l treatment of HIV-1 infected individuals. Mutations in the protease gene a re associated with reduced sensitivity to antiretroviral dugs. The aim of t his study was to detect mutations in HIV-1 protease gene from patients plas ma failing antiretroviral therapy with protease inhibitors through a sequen cing method The protease gene was amplified through RT-nested PCR. The ampl ification product was purified and dideoxy-sequencing on both strands. We h ave detected primary mutations, secondary mutations and unknown significanc e mutations in the protease gene of patients failing treatment regimen with protease inhibitors. For patients with drug-resistant HIV-1, results of re sistance testing may help guide the choice of salvage therapy. Thus, resist ance testing will be must useful in identifying drugs to be avoided. bur th e absence of apparent drug resistance is no guarantee of therapeutic succes s.