Methods for amplifying hepatitis C virus nucleic acids

US8679740B2 · kind B2 · utility

Assignee

• Vertex Pharmaceuticals Incorporated · US

Inventors

• Ann Dak-Yee Kwong · Cambridge, US
• James D. Frantz · Cape Coral, US
• Douglas John Bartels · North Liberty, US
• Chao Lin · Brookline, US
• Benjamin Shames · Arlington, US
• Sheila Seepersaud · Acton, US
• Judith A. Lippke · Sharon, US
• Tara Lynn Kieffer · Brookline, US
• Yi Zhou · Lexington, US
• Eileen Z. Zhang · Andover, US
• James C. Sullivan · Sudbury, US

Key dates

• Filing date: Jul 20, 2011
• Grant date: Mar 25, 2014
• Expiry date: Jul 20, 2031

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6883
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

A method of amplifying an HCV nucleic acid in an HCV infected sample comprises amplifying a segment of a DNA template that is complementary to a genome of HCV RNA from the sample by a two-stage PCR, wherein a first stage PCR employs a first outer primer and a second outer primer, and a second stage PCR employs a first inner primer and a second inner primer. The nucleotide sequence of the first outer primer comprises a nucleotide sequence as set forth in SEQ ID NO: 2; or SEQ ID NO:9, wherein optionally 1, 2 or 3 nucleotides are other nucleotides than those of SEQ ID NO: 9. The nucleotide sequence of the second outer primer comprises a nucleotide sequence set forth in SEQ ID NO: 3 or 4; or a nucleotide sequence as set forth in SEQ ID NO: 10 or 11, wherein optionally 1, 2 or 3 nucleotides are other nucleotides than those of SEQ ID NO: 10 and 11. The nucleotide sequence of the first inner primer comprises a nucleotide sequence as set forth in SEQ ID NO: 5; or SEQ ID NO:12, wherein optionally 1, 2 or 3 nucleotides are other nucleotides than those of SEQ ID NO: 12. The nucleotide sequence of the second inner primer comprises a nucleotide sequence as set forth in SEQ ID NO: 6 or 7; or a n…