Displacement polynucleotide assay employing polyether and diagnostic kit

US4766064A · kind A · utility

Assignees

• ALLIED CONSULTING, INC. · US
• Genetics Institute, LLC · US

Inventors

• Jon I. Williams · Lawrence, US
• Marian S. Ellwood · Lebanon, US
• Mary Collins · Natick, US
• Edward F. Fritsch · Concord, US
• Joseph G. Brewen · Madison, US
• Steven E. Diamond · Springfield, US

Key dates

• Filing date: Dec 20, 1984
• Grant date: Aug 23, 1988
• Expiry date: Dec 20, 2004

Classification

• Technology area (CPC Y): Emerging Cross-Sectional Technologies
• CPC primary: Y10S435/81
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

A diagnostic reagent is disclosed containing a complex of a probe polynucleotide (P) bound via purine/pyrimidine hydrogen bonding to a labeled polynucleotide (L). The probe (P) contains a target binding region (TBR) capable of binding to a target nucleotide sequence (G) of a biological sample. A method is disclosed in which contact with a sample containing the target nucleotide sequence (G) causes binding, initially between G and a single-stranded portion (IBR) of the target binding region (TBR). Thereafter the labeled polynucleotide (L) is displaced from the complex by branch migration of (G) into the (P)/(L) binding region. A volume excluding polymeric agent such as poly(ethylene oxide) (PEO or PEG) or other polyethers enhances the rate of appearance of displaced labeled polynucleotide. Determination of displaced labeled polynucleotide (L) gives a value which is a function of the presence and concentration of target nucleotide sequence (G) in the sample.