C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
150/3, 150/8.5
C12Q 1/68 (2006.01)
Patent
CA 1254114
ABSTRACT DISPLACEMENT POLYNUCLEOTIDE ASSAY EMPLOYING RECOMBINATION PROTEIN AND DIAGNOSTIC KIT A diagnostic reagent is disclosed containing a com- plex 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) in the presence of a recombination protein and its cofactors 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. Determination of displaced labeled polynucleo- tide (L) gives a value which is a function of the presence and concentration of target nucleotide sequence (G) in the sample. The presence of the recombination protein (such as the rec A protein from the enteric bacterium E. coli) enhances the rate of appearance of displaced labeled polynucleotide, especially when the reaction is conducted at or near physiological temperatures.
480559
Brewen Joseph G.
Collins Mary
Diamond Steven E.
Ellwood Marian S.
Fritsch Edward F.
Alliedsignal Inc.
Gowling Lafleur Henderson Llp
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