Primers and probes for the amplification and detection of HIV GAG, REV and NEF polynucleotides
The invention relates to improved methods and compositions for the nucleic acid amplification of one or multiple variants (strains) of Human Immunodeficiency Virus (HIV) present in a sample, and preferably in a sample from a pathogen infected individual. In particular, novel primers, methods and kits for the amplification of one or more species of HIV Rev, Gag and Nef nucleic acids are provided. The amplified HIV nucleic acid can be used to identify and/or quantitate HIV variants present in a sample. Nucleic acids produced by the methods of the invention or the proteins encoded thereby can also be used directly as vaccines or to transfect/load antigen presenting cells. The loaded antigen presenting cells can be used as a vaccine for the treatment or prevention of HIV infection.
1. A method for amplifying an HIV Gag nucleic acid, comprising carrying out a polymerase chain reaction using the Gag forward primer of SEQ ID NO:30 and the Gag reverse primers of SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47 and SEQ ID NO:48.
2. A method for amplifying an HIV Gag nucleic acid, comprising carrying out a polymerase chain reaction using the Gag forward primer of SEQ ID NO:31 and the Gag reverse primers SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47 and SEQ ID NO:48.
3. The method of claim 2 , further comprising using the Gag forward primer SEQ ID NO:30.