COMPOSITIONS FOR USE IN IDENTIFICATION OF STRAINS OF HEPATITIS C VIRUS
First Claim
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1. An oligonucleotide primer pair comprising a forward primer and a reverse primer, each between 13 and 35 linked nucleotides in length, said primer pair configured to generate an amplification product between 45 and 200 linked nucleotides in length, said forward primer configured to hybridize with at least 70% complementarity to a first portion of a region defined by nucleotide residues 9177 to 9337 of Genbank Accession Number:
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001433.1, and said reverse primer configured to hybridize with at least 70% complementarity to said second portion of said region.
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Abstract
The present invention provides compositions, kits and methods for rapid identification and quantification of strains of hepatitis C viruses by molecular mass and base composition analysis.
99 Citations
46 Claims
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1. An oligonucleotide primer pair comprising a forward primer and a reverse primer, each between 13 and 35 linked nucleotides in length, said primer pair configured to generate an amplification product between 45 and 200 linked nucleotides in length, said forward primer configured to hybridize with at least 70% complementarity to a first portion of a region defined by nucleotide residues 9177 to 9337 of Genbank Accession Number:
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001433.1, and said reverse primer configured to hybridize with at least 70% complementarity to said second portion of said region. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
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17. An oligonucleotide primer pair comprising a forward primer and a reverse primer, each between 13 and 35 linked nucleotides in length wherein said forward primer has at least 70% sequence identity with SEQ ID NO:
- 2 and said reverse primer has at least 70% sequence identity with SEQ ID NO;
29. - View Dependent Claims (18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30)
- 2 and said reverse primer has at least 70% sequence identity with SEQ ID NO;
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31. A kit for identifying a strain of hepatitis C virus, comprising:
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i) a first oligonucleotide primer pair comprising a forward primer and a reverse primer, each between 13 and 35 linked nucleotides in length, said primer pair configured to generate an amplification product that is between 45 and 200 linked nucleotides in length, said forward primer configured to hybridize with at least 70% complementarity to a first portion of a region defined by nucleotide residues 9177 to 9337 of Genbank Accession Number;
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001433.1, and said reverse primer configured to hybridize with at least 70% complementarity to a second portion of said region; andii) at least one additional primer pair, wherein the primers of each of said at least one additional primer pair are configured to hybridize to conserved sequence regions within genome segments of a hepatitis C genome, said genome segments selected from the group consisting of;
NS2, NS3 and NS5. - View Dependent Claims (32)
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33. A method for identifying a strain of hepatitis C virus in a sample, comprising:
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a) amplifying a nucleic acid from said sample using an oligonucleotide primer pair comprising a forward primer and a reverse primer, each between 13 and 35 linked nucleotides in length, said primer pair configured to generate an amplification product that is between 45 and 200 linked nucleotides in length, said forward primer configured to hybridize with at least 70% complementarity to a first portion of a region defined by nucleotide residues 9177 to 9337 of Genbank Accession Number;
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001433.1, and said reverse primer configured to hybridize with at least 70% complementarity to a second portion of said region;
wherein said amplifying step generates at least one amplification product that comprises between 45 and 200 linked nucleotides; andb) determining the molecular mass of said at least one amplification product by mass spectrometry. - View Dependent Claims (34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46)
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Specification