Reagents and methods for detecting HCV
AssigneeSiemens Healthcare Diagnostics Inc.
InventorsArejas Uzgiris, Sunil Pandit, Lance Palmer
Life of this patent
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Abstract
The present disclosure relates to oligonucleotide sequences for amplification primers and their use in performing nucleic acid amplifications of HCV, in particular regions that encode the NS3 polypeptide. In some embodiments the primers are used in nested PCR methods for the detection or sequencing of HCV NS3. The oligonucleotide sequences are also provided assembled as kits that can be used to amplify and detect or sequence HCV NS3.
Claims17
A method of performing a polymerase chain reaction (PCR) on an HCV NS3 1a target nucleic acid sequence in a sample comprising:
- contacting the sample with a set of outer primers, wherein the set of outer primers comprises one or more forward primers and one or more reverse primers,
- wherein the one or more forward primers comprise:
- a primer comprising SEQ ID NO:
- 1 or an active fragment thereof,
- a primer comprising SEQ ID NO:
- 2 or an active fragment thereof,
- a primer comprising SEQ ID NO:
- 3 or an active fragment thereof,
- a primer comprising SEQ ID NO:
- 4 or an active fragment thereof,
- a primer comprising SEQ ID NO:
- 5 or an active fragment thereof, or
- a combination thereof;
- and
- wherein the one or more reverse primers comprise:
- a primer comprising SEQ ID NO:
- 6 or an active fragment thereof,
- a primer comprising SEQ ID NO:
- 7 or an active fragment thereof,
- a primer comprising SEQ ID NO:
- 8 or an active fragment thereof,
- a primer comprising SEQ ID NO:
- 9 or an active fragment thereof,
- a primer comprising SEQ ID NO:
- 10 or an active fragment thereof, or
- a combination thereof;
- wherein an active fragment is 15 or more nucleotides in length;
- and submitting the resulting mixture to a first PCR.
A method of performing a polymerase chain reaction (PCR) on an HCV NS3 1a target nucleic acid sequence in a sample comprising:
- contacting the sample with a set of outer primers, wherein the set of outer primers comprises one or more forward primers and one or more reverse primers,
- wherein the one or more forward primers comprise one of:
- a primer comprising SEQ ID NO:
- 1,
- a primer comprising SEQ ID NO:
- 2,
- a primer comprising SEQ ID NO:
- 3,
- a primer comprising SEQ ID NO:
- 4, or
- a primer comprising SEQ ID NO:
- 5;
- and
- wherein the one or more reverse primers comprise one of:
- a primer comprising SEQ ID NO:
- 6,
- a primer comprising SEQ ID NO:
- 7,
- a primer comprising SEQ ID NO:
- 8,
- a primer comprising SEQ ID NO:
- 9, or
- a primer comprising SEQ ID NO:
- 10;
- and
- submitting the resulting mixture to a first PCR.
Dependencies are inferred from the claim text.