The invention relates to a test to detect mutations associated with antiretroviral drug resistance in HIV-1 using an allele-specific real-time PCR assay. The method uses mutation- specific PCR primers designed to detect and quantify the subset of HIV-1 viruses carrying the nucleic acid changes which are associated with cause resistance. The invention provides primers, primer sets, and amplification conditions which offer increased sensitivity of detection, after having designed and tested many different primers, primer sets, and PCR conditions. In addition, the method of the invention can be used to estimate the relative proportion of resistant viruses in the total virus population in a host. This method is therefore suited for the detection and quantification of enfuvirtide, protease, and reverse transcriptase resistance mutations in HIV-1 and will provide more useful clinical information for patient management.
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