Method of detecting genetic deletions identified with chromosomal abnormalities
First Claim
1. A method for identifying a chromosomal deletion, comprising the steps of:
- hybridizing a first probe set to a metaphase chromosome spread or an interphase nucleus, said first probe set being specific for-a deletion region of a chromosome but less than an entire chromosome such that said first probe set hybridizes to the deletion region if it is present within the metaphase chromosome spread or the interphase nucleus, and does not substantially hybridize to any region if the deletion region is absent from the metaphase chromosome spread or interphase nucleus, andhybridizing a second probe set to the metaphase chromosome spread or an interphase nucleus, said second probe set being specific for nucleotides in a region of the chromosome flanking the deletion region but less than an entire chromosome such that said second probe set will hybridize to the flanking region regardless of whether the deletion region is present or absent within the metaphase chromosome spread or interphase nucleus;
wherein (a) said first and second probe sets are distinct from one another and are each detectably labeled, (b) the label for the first probe set is distinguishable from the label for the second probe set and (c) each of the first and second probes has a complexity of at least 50,000 bases.
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Accused Products
Abstract
Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyzes. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods and reagents are provided for the detection of genetic rearrangements. Probes and test kits are provided for use in detecting genetic rearrangements, particularly for use in tumor cytogenetics, in the detection of disease related loci, specifically cancer, such as chronic myelogenous leukemia (CML) and for biological dosimetry. Methods and reagents are described for cytogenetic research, for the differentiation of cytogenetically similar but genetically different diseases, and for many prognostic and diagnostic applications.
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Citations
11 Claims
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1. A method for identifying a chromosomal deletion, comprising the steps of:
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hybridizing a first probe set to a metaphase chromosome spread or an interphase nucleus, said first probe set being specific for-a deletion region of a chromosome but less than an entire chromosome such that said first probe set hybridizes to the deletion region if it is present within the metaphase chromosome spread or the interphase nucleus, and does not substantially hybridize to any region if the deletion region is absent from the metaphase chromosome spread or interphase nucleus, and hybridizing a second probe set to the metaphase chromosome spread or an interphase nucleus, said second probe set being specific for nucleotides in a region of the chromosome flanking the deletion region but less than an entire chromosome such that said second probe set will hybridize to the flanking region regardless of whether the deletion region is present or absent within the metaphase chromosome spread or interphase nucleus; wherein (a) said first and second probe sets are distinct from one another and are each detectably labeled, (b) the label for the first probe set is distinguishable from the label for the second probe set and (c) each of the first and second probes has a complexity of at least 50,000 bases. - View Dependent Claims (2, 3, 8)
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4. A method of genotyping a sample, comprising:
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providing first and second chromosome-specific probes, each probe having a complexity of at least 50,000 bases; hybridizing the first and second chromosome-specific probes to the sample, the hybridizing resulting in sample staining with a contrast ratio of at least 4; and counting, in an interphase nucleus in the sample, a number of bands for the first and second chromosome-specific probes, wherein a first number of bands indicates that the sample has a first genotype and a second number of bands indicates that the sample has a second genotype. - View Dependent Claims (5, 6, 7)
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9. A method for identifying a chromosomal deletion, comprising the steps of:
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hybridizing a first probe set to a metaphase chromosome spread or an interphase nucleus, said first probe set being sufficient in length and substantially complementary to a deletion region of a chromosome but less than an entire chromosome such that said first probe set hybridizes to the deletion region if it is present within the metaphase chromosome spread or the interphase nucleus, and does not substantially hybridize to any region if the deletion region is absent from the metaphase chromosome spread or interphase nucleus; and hybridizing a second probe set to the metaphase chromosome spread or an interphase nucleus, said second probe set being sufficient in length and substantially complementary to nucleotides in a region of the chromosome flanking the deletion region but less than an entire chromosome such that said second probe set will hybridize to the flanking region regardless of whether the deletion region is present or absent within the metaphase chromosome spread or interphase nucleus; wherein (a) said first and second probe sets are distinct from one another and are each detectably labeled, (b) the label for the first probe set is distinguishable from the label for the second probe set and (c) each of the first and second probes has a complexity of at least 50,000 bases. - View Dependent Claims (10, 11)
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Specification