Polymerase chain reaction assays for monitoring antiviral therapy and making therapeutic decisions in the treatment of acouired immunodeficiency syndrome

Abstract

The present invention relates to methods of monitoring, via polymerase chain reaction, the clinical progression of human immunodeficiency virus infection and its response to antiretroviral therapy. According to the invention, polymerase chain reaction assays may be used to predict immunological decline and to identify, at an early stage, patients whose infection has become resistant to a particular antiretroviral drug regimen.

1 . A kit for determining the presence of a mutation at a codon of interest of the HIV reverse transcriptase enzyme, comprising a first pair of polymerase chain reaction (PCR) primers for amplifying by PCR a DNA fragment containing the codon of interest, and at least two pairs of second primers for amplifying DNA encoding wild type and mutant forms at the codon of interest.  
     
     
         2 . The kit of  claim 1  wherein the codon of interest is codon 215.  
     
     
         3 . The kit of  claim 1  wherein the codon of interest is codon 74.  
     
     
         4 . The kit of  claim 1  wherein the two pairs of second primers comprise primer B (5′-GGATGGAAAGGATCACC-3′) (SEQ ID NO: 3) and 3M (3′-AAGTGTGGTCTGTTTTTTGTA-5′)(SEQ ID NO: 5), and primers B and 3W (3′-TGGTGTGGTCTGTTTTTTGTA-5) (SEQ ID NO: 4).