Composition and method for the detection of the presence of a polynucleotide sequence of interest
First Claim
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1. In a composition for detecting the presence of a polynucleotide sequence of interest in a sample of polynucleotide sequences comprising:
- a composition comprising at least one first and at least one second polynucleotide the improvements comprising;
(a) said first and second sequences are labeled with a first detectable marker and are either present as separate molecules from which said first polynucleotide sequence has not been isolated or are covalently linked and further characterized in that said first sequence is capable of hybridizing to said sequence of interest and said second sequence is not capable of hybridizing to said sequence of interest and is not capable of hybridizing to said first sequence; and
(b) a composition comprising at least one third polynucleotide sequence that is unlabeled or is labeled with a second detectable marker and characterized in that said third sequence is not capable of hybridizing to said first sequence and is not capable of hybridizing to said sequence of interest, and further characterized in that said third sequence is;
(i) capable of hybridizing to said second sequence, such that by hybridizing to said second sequence said third sequence blocks hybridization between said second sequence and complementary non-target sequences that may be contained in the sample and decreased the likelihood that said second sequence will generate a false positive signal upon detection of said first detectable marker;
or(b) substantially identical to said second sequence, such that by means of hybridizing to said complementary non-target sequences that may be contained in the sample said third sequence blocks hybridization between said second sequence and said complementary non-target sequences and decreases the likelihood that said second sequence will generate a false positive signal upon detection of said first detectable marker.
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Abstract
The present invention relates to a wide range of genetic analyses using the technique of nucleic acid hybridization. These genetic analyses include, for example, the diagnosis of infections by foreign microbes and the detection of specific genetic traits and abnormalities. More specifically, the present invention is related to the detection of the presence of a polynucleotide sequence of interest.
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Citations
22 Claims
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1. In a composition for detecting the presence of a polynucleotide sequence of interest in a sample of polynucleotide sequences comprising:
a composition comprising at least one first and at least one second polynucleotide the improvements comprising; (a) said first and second sequences are labeled with a first detectable marker and are either present as separate molecules from which said first polynucleotide sequence has not been isolated or are covalently linked and further characterized in that said first sequence is capable of hybridizing to said sequence of interest and said second sequence is not capable of hybridizing to said sequence of interest and is not capable of hybridizing to said first sequence; and (b) a composition comprising at least one third polynucleotide sequence that is unlabeled or is labeled with a second detectable marker and characterized in that said third sequence is not capable of hybridizing to said first sequence and is not capable of hybridizing to said sequence of interest, and further characterized in that said third sequence is; (i) capable of hybridizing to said second sequence, such that by hybridizing to said second sequence said third sequence blocks hybridization between said second sequence and complementary non-target sequences that may be contained in the sample and decreased the likelihood that said second sequence will generate a false positive signal upon detection of said first detectable marker;
or(b) substantially identical to said second sequence, such that by means of hybridizing to said complementary non-target sequences that may be contained in the sample said third sequence blocks hybridization between said second sequence and said complementary non-target sequences and decreases the likelihood that said second sequence will generate a false positive signal upon detection of said first detectable marker. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
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12. In a method for the detection of a polynucleotide sequence of interest in a sample of polynucleotide sequences comprising:
contacting under hybridizing conditions said sample of polynucleotide sequences with a composition comprising at least one first and at least one second polynucleotide and detecting said polynucleotide of interest by means of said first detectable marker, the improvements comprising; (a) said first and second sequences are labeled with a first detectable marker and are either present as separate molecules from which said first polynucleotide sequence has not been isolated or are covalently linked and further characterized in that said first sequence is capable of hybridizing to said sequence of interest and said second sequence is not capable of hybridizing to said sequence of interest and is not capable of hybridizing to said first sequence; and (b) said sample is further contacted, prior to detecting, with a composition comprising at least one-third polynucleotide sequence that is unlabeled or is labeled with a second detectable marker and characterized in that said third sequence is not capable of hybridizing to said first sequence and is not capable of hybridizing to said sequence of interest, and further characterized in that said third sequence is; (i) capable of hybridizing to said second sequence, such that by hybridizing to said second sequence said third sequence blocks hybridization between said second sequence and complementary non-target sequences that may be contained in the sample and decreases the likelihood that said second sequence will generate a false positive signal upon detection of said first detectable marker;
or(ii) substantially identical to said second sequence, such that by means of hybridizing to said complementary non-target sequences that may be contained in the sample said third sequence blocks hybridization between said second sequence and said complementary non-target sequences and decreases the likelihood that said second sequence will generate a false positive signal upon detection of said first detectable marker. - View Dependent Claims (13, 14, 15, 16, 17, 18, 19, 20, 21, 22)
Specification