Probes and methods for detection of nucleic acids
AssigneeBecton, Dickinson and Company
InventorsJames G. Nadeau, Tobin J. Hellyer
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Abstract
The invention employs an unlabeled signal primer comprising a 5′ adapter sequence for detection of nucleic acid target sequences. The detection system further comprises a reporter probe, the 3′ end of which hybridizes to the complement of the 5′ adapter sequence of the signal primer to produce a 5′ overhang. Polymerase is used to fill in the overhang and synthesize the complement of the 5′ overhang of the reporter probe. Synthesis of the reporter probe complement is detected, either directly or indirectly, as an indication of the presence of the target.
Claims15
A method for detecting amplification of a target sequence comprising, in an amplification reaction:
- a) hybridizing a signal primer comprising an adapter sequence to the target sequence;
- b) extending the signal primer on the target sequence to produce an extension product;
- c) separating the extension product from the target sequence;
- d) hybridizing an amplification primer to the extension product and extending the amplification primer to synthesize a complement of the adapter sequence;
- e) separating the product including the complement of the adapter sequence from the extension product;
- f) hybridizing to the complement of the adapter sequence a reporter probe comprising a reporter moiety, and producing a double-stranded reporter moiety;
- and
- g) detecting the double-stranded reporter moiety as an indication of amplification of the target sequence.
Dependencies are inferred from the claim text.