Oligonucleotide probes and primers comprising universal bases for diagnostic purposes
First Claim
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1. An oligonucleotide probe for detecting a first target nucleic acid sequence comprising the formula:
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XRYwherein X comprises between about 5 and 20 modified nucleic acid bases complementary to a first region of said first target nucleic acid sequence;
wherein R comprises between about 5 and 10 universal or generic bases juxtaposed to one another and said universal or generic bases align with bases of a second region of said first target nucleic acid sequence;
wherein Y comprises between about 5 and about 20 nucleic acid bases complementary to a third region of said first target nucleic acid sequence;
wherein X, R, and Y are covalently joined;
wherein greater than 20% of the total number of bases in said oligonucleotide probe are said universal or generic bases juxtaposed to one another;
wherein said oligonucleotide probe further comprises a detectable label; and
wherein said oligonucleotide probe is capable of hybridizing to and detecting said first target nucleic acid sequence but not able to detect a second target nucleic acid sequence that differs from said first target nucleic acid sequence by a single nucleotide base mismatch in the first or third region of the target nucleic acid sequence.
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Abstract
Aspects of the invention relate to novel oligonucleotides comprising universal and generic bases for use as primers and probes, as well as, methods of using said oligonucleotides for the diagnosis of disease.
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23 Claims
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1. An oligonucleotide probe for detecting a first target nucleic acid sequence comprising the formula:
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XRYwherein X comprises between about 5 and 20 modified nucleic acid bases complementary to a first region of said first target nucleic acid sequence; wherein R comprises between about 5 and 10 universal or generic bases juxtaposed to one another and said universal or generic bases align with bases of a second region of said first target nucleic acid sequence; wherein Y comprises between about 5 and about 20 nucleic acid bases complementary to a third region of said first target nucleic acid sequence; wherein X, R, and Y are covalently joined; wherein greater than 20% of the total number of bases in said oligonucleotide probe are said universal or generic bases juxtaposed to one another; wherein said oligonucleotide probe further comprises a detectable label; and wherein said oligonucleotide probe is capable of hybridizing to and detecting said first target nucleic acid sequence but not able to detect a second target nucleic acid sequence that differs from said first target nucleic acid sequence by a single nucleotide base mismatch in the first or third region of the target nucleic acid sequence. - View Dependent Claims (3, 4, 5, 6, 7, 8, 9, 10, 11, 21, 22, 23)
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2. An oligonucleotide probe for distinguishing a first target nucleic acid sequence from a second target nucleic acid sequence that differs from said first target nucleic acid sequence by a nucleotide base mismatch, said oligonucleotide probe comprising
a site complementary to said first target nucleic acid sequence, said site having a 5′ - and 3′
side thereto and having a non-complementary base to said mismatch of said second target nucleic acid sequence;at least five universal or generic bases juxtaposed to one another, said universal or generic bases being located on the 5′
or 3′
side of said site and aligned with bases of the first target nucleic acid sequence;a first region complementary to a first region of the first target nucleic acid and a second region complementary to a second region of the first target nucleic acid, said first and second regions being located on the 5′ and
3′
side of said site, respectively;wherein said oligonucleotide probe has between about 10 and 25 bases; wherein said oligonucleotide probe further comprises a detectable label; and wherein said probe detects said first target nucleic acid sequence and does not detect said second target sequence. - View Dependent Claims (12, 13, 14, 15, 16, 17, 18, 19, 20)
- and 3′
Specification