PCR method

US7101663B2 · kind B2 · utility

Assignee

• University of Pittsburgh—Of the Commonwealth System of Higher Education · US

Inventors

• Tony Godfrey · San Francisco, US
• James Luketich · Pittsburgh, US
• Siva Raja · Pittsburgh, US
• Lori Kelly · Nazareth, US
• Sydney David Finkelstein · Pittsburgh, US

Key dates

• Filing date: Mar 4, 2002
• Grant date: Sep 5, 2006
• Expiry date: Aug 5, 2023

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q2600/166
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

A method for balancing multiplexed PCR methods is provided. In the method, two or more sequential temporal PCR stages are used to effectively separate two or more PCR reactions in a single tube as an alternative to primer limiting to modulate the relative rate of production of a first amplicon by a first primer set and a second amplicon by a second primer set during the first and second amplification stages. Also provided are rapid RT-PCR methods that find particular use in intraoperative diagnoses and prognoses, for instance in diagnosing malignant esophageal adenocarcenoma by determining expression levels of carcinoembryonic antigen (CEA) in sentinel lymph nodes.