Strand-invasion based dna amplification method
AssigneeOrion Diagnostica Oy
InventorSanna FILÉN
Life of this patent
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Abstract
A method for detecting toxigenic Clostridium difficile (C. difficile) by strand-invasion based DNA amplification is provided, together with oligonucleotides, compositions and kits suitable for use in this method.
Claims18
A method for detecting a target nucleic acid sequence of toxigenic C. difficile in a sample, said method comprising contacting said sample with at least one upstream primer, at least one downstream primer and at least one strand invasion oligonucleotide under conditions promoting amplification of said target nucleic acid sequence,
- wherein each said primer and said strand invasion oligonucleotide comprises a region complementary to said target nucleic acid sequence;
- and
- wherein said strand invasion oligonucleotide renders at least a portion of the target nucleic acid sequence single-stranded to allow the binding of said upstream primer and a downstream primer.
A kit comprising at least two oligonucleotides selected from (a) an upstream primer, (b) a downstream primer and (c) a strand invasion oligonucleotide,
- wherein each oligonucleotide-comprises-one or more modified nucleotides in its 3′ region—and wherein said upstream primer is an oligonucleotide of fewer than 30 nucleotides in length comprising the sequence of SEQ ID NO:
- 2, SEQ ID NO:7, or a variant thereof, and/or wherein said downstream primer is an oligonucleotide of fewer than 30 nucleotides in length comprising the sequence of SEQ ID NO:
- 3, SEQ ID NO:8, or a variant thereof, and/or wherein said strand invasion oligonucleotide is an oligonucleotide of greater than 30 nucleotides in length comprising the sequence of SEQ ID NO:
- 4, or SEQ ID NO:9, or a variant thereof.
(canceled)
(canceled)
A composition comprising at least two oligonucleotides selected from (a) an upstream primer, (b) a downstream primer and (c) a strand invasion oligonucleotide,
- wherein each-oligonucleotide-comprises-one or more modified nucleotides in its 3′region—and wherein—said upstream primer is an oligonucleotide of fewer than 30 nucleotides in length comprising the sequence of SEQ ID NO:
- 2, SEQ ID NO:7, or a variant thereof, and/or wherein said downstream primer is an oligonucleotide of fewer than 30 nucleotides in length comprising the sequence of SEQ ID NO:
- 3, SEQ ID NO:8, or a variant thereof, and/or wherein said strand invasion oligonucleotide is an oligonucleotide of greater than 30 nucleotides in length comprising the sequence of SEQ ID NO:
- 4, or SEQ ID NO:9, or a variant thereof.
Dependencies are inferred from the claim text.