Patent EP0364255A2 successfully refreshed from Google Patents.
EP0364255A2
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Multiplex genomic DNA amplification for deletion detection
Assignee: Baylor College of Medicine
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Filing: 1989-10-11
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Granted: —
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Expires: 2009-10-11
Abstract
Abstract The present invention relates to a method for detecting multiple DNA sequences simultaneously. The method involves amplification of multiple sequences simultaneously by annealing a plurality of paired oligonucleotide primers to single stranded DNA. One member of each pair is complimentary to the sense strand of a sequences and the other member is complimentary to a different segment of the anti-sense strand of the same sequence. The amplification occurs by alternately annealing and extending the primers. The invention also includes oligonucleotide primer sequences helpful in detecting genetic diseases and/or exogenous DNA sequences.
Inventors
- 👤 Charles T. Caskey
- 👤 Jeffrey S. Chamberlain
- 👤 Richard A.L. Gibbs
- 👤 Joel E. Rainer
- 👤 Phi Nga Nguyen
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-31 07:55
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6876; C12Q1/6883; C07; C07K; C07K14/00; C07K14/435; C07K14/46; C07K14/47; C07K14/4701; C07K14/4707; C07K14/4708; C12Q1/6844; C12Q1/6858; C12Q1/686; C12Q2600/00; C12Q2600/156; C12Q2600/16
Claims Analysis
Total Claims: 27
Claim 1
Independent Claim
172 words
A method for simultaneously detecting deletions at a plurality of DNA sequences, comprising the steps of: treating said DNA to form single-stranded complementary strands; adding a plurality of paired oligonucleotide primers, each pair specific for a different sequence, one primer of each pair substantially complementary to a part of the sequence in the sense-strand and the other primer of each pair substantially complementary to a different part of the same sequence in the complementary anti-sense strand; annealing the plurality of primers to their complementary sequences; simultaneously extending said plurality of annealed primers from each primer's 3′ terminus to synthesize an extension product complementary to the strands annealed to each primer, said extension products, after separation from their complement, serving as templates for the synthesis of an extension product from the other primer of each pair; separating said extension products from said templates to produce single-stranded molecules; amplifying said single stranded molecules by repeating, at least once, said annealing, extending and separating steps; and identifying said amplified extension products from each different sequence.
Claim 2
Independent Claim
172 words
A method for simultaneously detecting deletions at a plurality of DNA sequences, comprising the steps of: treating said DNA to form single-stranded complementary strands; adding a plurality of paired oligonucleotide primers, each pair specific for a different sequence, one primer of each pair substantially complementary to a part of the sequence in the sense-strand and the other primer of each pair substantially complementary to a different part of the same sequence in the complementary anti-sense strand; annealing the plurality of primers to their complementary sequences; simultaneously extending said plurality of annealed primers from each primer's 3′ terminus to synthesize an extension product complementary to the strands annealed to each primer, said extension products, after separation from their complement, serving as templates for the synthesis of an extension product from the other primer of each pair; separating said extension products from said templates to produce single-stranded molecules; amplifying said single stranded molecules by repeating, at least once, said annealing, extending and separating steps; and identifying said amplified extension products from each different sequence.
Claim 3
Dependent Claim
31 words
The method of Claim 1 for detecting deletions at a plurality of genomic DNA sequences, wherein said sequences are selected from the group of sequences on the X and Y chromosomes.
Claim 4
Dependent Claim
24 words
The method of Claim 2 for the detection of X-linked disease, wherein said genomic DNA sequences contain a deletion that causes a genetic disease.
Claim 5
Dependent Claim
32 words
The method of Claim 3 for the detection of said X-linked genetic diseases selected from the group consisting of ornithine transcarbamylase deficiency, hypoxanthine phosphoribosyltransferfase deficiency, steroid sulfatase deficiency and X-linked muscular dystrophy.
Claim 6
Dependent Claim
31 words
The method of Claim 4 for the detection of X-linked muscular dystrophy, wherein said plurality of paired primers are complementary to different sequences within the gene coding for the dystrophin protein.
Claim 7
Dependent Claim
47 words
The method of Claim 5, wherein the plurality of paired primers is selected from the group consisting of: (1) 5′-GACTTTCGATGTTGAGATTACTTTCCC-3′ (2) 5′-AAGCTTGAGATGCTCTCACCTTTTCC-3′, (1) 5′-GTCCTTTACACACTTTACCTGTTGAG-3′ (2) 5′-GGCCTCATTCTCATGTTCTAATTAG-3′, (1) 5′-AAACATGGAACATCCTTGTGGGGAC-3′ (2) 5′-CATTCCTATTAGATCTGTCGCCCTAC-3′, (1) 5′-GATAGTGGGCTTTACTTACATCCTTC-3′ (2) 5′-GAAAGCACGCAACATAAGATACACCT-3′, (1) 5′-CTTGATCCATATGCTTTTACCTGCA-3′ (2) 5′-TCCATCACCCTTCAGAACCTGATCT-3′, (1) 5′-GAATACATTGGTTAAATCCCAACATG-3′ (2) 5′-CCTGAATAAAGTCTTCCTTACCACAC-3′, and (1) 5′-TTCTACCACATCCCATTTTCTTCCA-3′ (2) 5′-GATGGCAAAAGTGTTGAGAAAAAGTC-3′.
Claim 8
Dependent Claim
13 words
The method of Claim 3, wherein said genomic DNA is from fetal tissue.
Claim 9
Dependent Claim
70 words
The method of Claim 1 for detecting deletions at a plurality of genomic DNA sequences, wherein the plurality of paired primers is selected from the group consisting of: (1) 5′-GACTTTCGATGTTGAGATTACTTTCCC-3′ (2) 5′-AAGCTTGAGATGCTCTCACCTTTTCC-3′, (1) 5′-GTCCTTTACACACTTTACCTGTTGAG-3′ (2) 5′-GGCCTCATTCTCATGTTCTAATTAG-3′, (1) 5′-AAACATGGAACATCCTTGTGGGGAC-3′ (2) 5′-CATTCCTATTAGATCTGTCGCCCTAC-3′, (1) 5′-GATAGTGGGCTTTACTTACATCCTTC-3′ (2) 5 -GAAAGCACGCAACATAAGATACACCT-3′, (1) 5′-CTTGATCCATATGCTTTTACCTGCA-3′ (2) 5′-TCCATCACCCTTCAGAACCTGATCT-3′, (1) 5′-GAATACATTGGTTAAATCCCAACATG-3′ (2) 5′-CCTGAATAAAGTCTTCCTTACCACAC-3′, (1) 5′-TTCTACCACATCCCATTTTCTTCCA-3′ (2) 5′-GATGGCAAAAGTGTTGAGAAAAAGTC-3′, (1) 5′-TGGTCTCCTTAAACCTGTCTT-3′ (2) 5′-ACACAACTGTGTTCACTAG-3′, (1) 5′-ACGTGGAGTGACGATGCTCTTCCC-3′ (2) 5′-GTGGGATTCACCACTTTTCCC-3′, and (1) 5′-GAAGTCAAGGACACCGAGGAA-3′ (2) 5′-AGCCCTCTGGCCAGTCCTAGTG-3′.
Claim 10
Dependent Claim
46 words
A DNA sequence of the formula: and fragments and derivatives thereof, said fragments and derivatives complementary to the sense and anti-sense strands of the gene coding for dystrophin, said fragments and derivatives capable of annealing to said strands of the dystrophin gene and amplifying dystrophin sequences.
Claim 11
Dependent Claim
46 words
A DNA sequence of the formula: and fragments and derivatives thereof, said fragments and derivatives complementary to the sense and anti-sense strands of the gene coding for dystrophin, said fragments and derivatives capable of annealing to said strands of the dystrophin gene and amplifying dystrophin sequences.
Claim 12
Dependent Claim
46 words
A DNA sequence of the formula: and fragments and derivatives thereof, said fragments and derivatives complementary to the sense and anti-sense strands of the gene coding for dystrophin, said fragments and derivatives capable of annealing to said strands of the dystrophin gene and amplifying dystrophin sequences.
Claim 13
Dependent Claim
46 words
A DNA sequence of the formula: and fragments and derivatives thereof, said fragments and derivatives complementary to the sense and anti-sense strands of the gene coding for dystrophin, said fragments and derivatives capable of annealing to said strands of the dystrophin gene and amplifying dystrophin sequences.
Claim 14
Dependent Claim
46 words
A DNA sequence of the formula: and fragments and derivatives thereof, said fragments and derivatives complementary to the sense and anti-sense strands of the gene coding for dystrophin, said fragments and derivatives capable of annealing to said strands of the dystrophin gene and amplifying dystrophin sequences.
Claim 15
Dependent Claim
46 words
A DNA sequence of the formula: and fragments and derivatives thereof, said fragments and derivatives complementary to the sense and anti-sense strands of the gene coding for dystrophin, said fragments and derivatives capable of annealing to said strands of the dystrophin gene and amplifying dystrophin sequences.
Claim 16
Dependent Claim
46 words
A DNA sequence of the formula: and fragments and derivatives thereof, said fragments and derivatives complementary to the sense and anti-sense strands of the gene coding for dystrophin, said fragments and derivatives capable of annealing to said strands of the dystrophin gene and amplifying dystrophin sequences.
Claim 17
Dependent Claim
46 words
A DNA sequence of the formula: and fragments and derivatives thereof, said fragments and derivatives complementary to the sense and anti-sense strands of the gene coding for dystrophin, said fragments and derivatives capable of annealing to said strands of the dystrophin gene and amplifying dystrophin sequences.
Claim 18
Dependent Claim
46 words
A DNA sequence of the formula: and fragments and derivatives thereof, said fragments and derivatives complementary to the sense and anti-sense strands of the gene coding for dystrophin, said fragments and derivatives capable of annealing to said strands of the dystrophin gene and amplifying dystrophin sequences.
Claim 19
Dependent Claim
46 words
A DNA sequence of the formula: and fragments and derivatives thereof, said fragments and derivatives complementary to the sense and anti-sense strands of the gene coding for dystrophin, said fragments and derivatives capable of annealing to said strands of the dystrophin gene and amplifying dystrophin sequences.
Claim 20
Dependent Claim
46 words
A DNA sequence of the formula: and fragments and derivatives thereof, said fragments and derivatives complementary to the sense and anti-sense strands of the gene coding for dystrophin, said fragments and derivatives capable of annealing to said strands of the dystrophin gene and amplifying dystrophin sequences.
Claim 21
Dependent Claim
46 words
A DNA sequence of the formula: and fragments and derivatives thereof, said fragments and derivatives complementary to the sense and anti-sense strands of the gene coding for dystrophin, said fragments and derivatives capable of annealing to said strands of the dystrophin gene and amplifying dystrophin sequences.
Claim 22
Dependent Claim
46 words
A DNA sequence of the formula: and fragments and derivatives thereof, said fragments and derivatives complementary to the sense and anti-sense strands of the gene coding for dystrophin, said fragments and derivatives capable of annealing to said strands of the dystrophin gene and amplifying dystrophin sequences.
Claim 23
Dependent Claim
46 words
A DNA sequence of the formula: and fragments and derivatives thereof, said fragments and derivatives complementary to the sense and anti-sense strands of the gene coding for dystrophin, said fragments and derivatives capable of annealing to said strands of the dystrophin gene and amplifying dystrophin sequences.
Claim 24
Dependent Claim
46 words
A DNA sequence of the formula: and fragments and derivatives thereof, said fragments and derivatives complementary to the sense and anti-sense strands of the gene coding for dystrophin, said fragments and derivatives capable of annealing to said strands of the dystrophin gene and amplifying dystrophin sequences.
Claim 25
Dependent Claim
46 words
A DNA sequence of the formula: and fragments and derivatives thereof, said fragments and derivatives complementary to the sense and anti-sense strands of the gene coding for dystrophin, said fragments and derivatives capable of annealing to said strands of the dystrophin gene and amplifying dystrophin sequences.
Claim 26
Dependent Claim
46 words
A DNA sequence of the formula: and fragments and derivatives thereof, said fragments and derivatives complementary to the sense and anti-sense strands of the gene coding for dystrophin, said fragments and derivatives capable of annealing to said strands of the dystrophin gene and amplifying dystrophin sequences.
Claim 27
Dependent Claim
46 words
A DNA sequence of the formula: and fragments and derivatives thereof, said fragments and derivatives complementary to the sense and anti-sense strands of the gene coding for dystrophin, said fragments and derivatives capable of annealing to said strands of the dystrophin gene and amplifying dystrophin sequences.
References Cited (Backward Citations)
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