Amplification of HIV-1 sequences for detection of sequences associated with drug-resistance mutations
AssigneesGen-Probe Incorporated, Biomerieux
InventorsYeasing Y. Yang, Steven T. Brentano, Odile Babola, Nathalie Tran, Guy Vernet
Life of this patent
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Abstract
Sequences of nucleic acid oligonucleotides for amplifying different portions of gag and pol genes of HIV-1 and for detecting such amplified nucleic acid sequences are disclosed. Methods of amplifying and detecting HIV-1 nucleic acid in a biological sample using the amplification oligonucleotides specific for gag and pol target sequences are disclosed.
Claims21
A mixture of nucleic acid oligomers for specifically amplifying a plurality of nucleotide sequences in distinct regions of the HIV-1 genome in pol and gag gene sequences, comprising:
- oligomers specific for a region in the pol sequence, referred to as a RT1 sequence, that include an oligonucleotide in a size range of 18 to 23 nucleotides that contains the sequence consisting of SEQ ID NO:14 or an oligonucleotide that contains a 5′ promoter sequence covalently attached to a sequence in a size range of 18 to 23 nucleotides that contains the sequence consisting of SEQ ID NO:14, and an oligonucleotide in a size range of 19 to 20 nucleotides that contains the sequence consisting of SEQ ID NO:64,
- oligomers specific for a region in the pol sequence, referred to as a RT4 sequence, that include an oligonucleotide consisting of SEQ ID NO:16 or an oligonucleotide that contains a 5′ promoter sequence covalently attached to the sequence consisting of SEQ ID NO:16, and an oligonucleotide consisting of SEQ ID NO:22, and
- oligomers specific for a region in the gag sequence, referred to as a Gag2 sequence, that include an oligonucleotide consisting of SEQ ID NO:12 or an oligonucleotide that contains a 5′ promoter sequence covalently attached to the sequence consisting of SEQ ID NO:12, and an oligonucleotide consisting of SEQ ID NO:18.
A method of detecting HIV-1 in a biological sample, comprising the steps of:
- providing a biological sample containing HIV-1 nucleic acid;
- mixing the sample with a mixture of amplification oligomers that specifically amplify a plurality of distinct regions in HIV-1 target sequences contained within a pol gene sequence, referred to as a RT1 sequence and a RT4 sequence, and contained in a gag sequence, referred to as a Gag2 sequence, under conditions that allow amplification of nucleic acid, wherein the amplification oligomers comprise
- oligomers specific for the RT1 sequence that include an oligonucleotide in a size range of 18 to 23 nucleotides that contains the sequence consisting of SEQ ID NO:14 or an oligonucleotide that contains a 5′ promoter sequence covalently attached to a sequence in a size range of 18 to 23 nucleotides that contains the sequence consisting of SEQ ID NO:14, and an oligonucleotide in a size range of 19 to 20 nucleotides that contains the sequence consisting of SEQ ID NO:64,
- oligomers specific for the RT4 sequence that include an oligonucleotide consisting of SEQ ID NO:16 or an oligonucleotide that contains a 5′ promoter sequence covalently attached to the sequence consisting of SEQ ID NO:16, and an oligonucleotide consisting of SEQ ID NO:22, and
- oligomers specific for the Gag2 sequence that include an oligonucleotide consisting of SEQ ID NO:12 ;
- or an oligonucleotide that contains a 5′ promoter sequence covalently attached to the sequence consisting of SEQ ID NO:12, and an oligonucleotide consisting of SEQ ID NO:18 to permit amplification of the RT1, RT4 and Gag2 sequences in the HIV-1 nucleic acid to produce amplified products of the HIV-1 nucleic acid;
- and
- detecting the presence of the amplified products of the HIV-1 nucleic acid to indicate the presence of HIV-1 in the biological sample.
Dependencies are inferred from the claim text.