Methods and compositions for chromosome 21-specific staining

• US 6,500,612 B1
• Filed: 06/07/1995
• Issued: 12/31/2002
• Est. Priority Date: 01/16/1986
• Status: Expired due to Term

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Pinkel, Daniel, Gray, Joe W.
• Primary Examiner(s): Marschel, Ardin H.
• Application Number:
  US08/472,312
• Time in Patent Office: 2,764 Days
• Field of Search: 435/6, 435/810, 436/501, 536/23.1, 536/24.1, 536/24-- 2, 536/25.3, 935/77.78
• US Class Current: 435/6.11
• CPC Class Codes: C12Q 1/6811 Selection methods for production or design of target specific oligonucleotides or binding molecules
• CPC Class Codes: C12Q 1/6832 Enhancement of hybridisation reaction
• CPC Class Codes: C12Q 1/6841 In situ hybridisation
• CPC Class Codes: C12Q 1/6853 using modified primers or templates
• CPC Class Codes: C12Q 1/6879 for sex determination
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2525/151 repeat or repeated sequences, e.g. VNTR, microsatellite, concatemer
• CPC Class Codes: C12Q 2525/186 incorporating a non-extendable or blocking moiety
• CPC Class Codes: C12Q 2563/107 fluorescence
• CPC Class Codes: C12Q 2563/131 the label being a member of a cognate binding pair, i.e. extends to antibodies, haptens, avidin
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 435/82 Subcellular parts of microorganisms

Abstract

Methods and compositions for chromosome-specific staining are provided. Composition comprise heterogenous mixtures of labeled nucleic acid fragments having substantially complementary base sequences to unique sequence regions of the chromosomal DNA for which their associated staining reagent is specific. Methods include methods for making the chromosome-specific staining compositions of the invention, and methods for applying the staining compositions to chromosomes.

Claims

1. A method of staining interphase chromosome 21 DNA, comprising:

(a) providing a chromosome 21-specific staining composition which comprises a heterogenous mixture of labeled unique sequence DNA fragments wherein said fragments are substantially complementary to DNA segments within human chromosome 21 DNA; and

(b) employing the heterogenous mixture and interphase chromosomal DNA in in situ hybridization to permit detection of labeled unique sequence DNA fragments which are hybridized to human chromosome 21 DNA, wherein the interphase chromosomal DNA is present in a morphologically identifiable cell nucleus during the in situ hybridization.

2. The method of claim 1 wherein the labeled DNA comprises fragments which are designed to allow detection of an extra chromosome 21.

3. The method of claim 1 wherein the labeled DNA comprises fragments which are designed to allow detection of a missing chromosome 21.

4. The method of claim 1 wherein the labeled DNA comprises fragments which are designed to allow detection of an extra portion of chromosome 21.

5. The method of claim 4 wherein the chromosomal rearrangement is a translocation.

6. The method of claim 4 wherein the chromosomal rearrangement is an inversion.

7. The method of claim 1 wherein the labeled DNA comprises fragments which are designed to allow detection of a missing portion of chromosome 21.

8. The method of claim 1 wherein the labeled DNA comprises fragments designed to allow detection of a chromosomal rearrangement of chromosome 21.

9. The method of claim 1 further comprising labeling total DNA in said cell nucleus with a second label that is distinguishable from the label used to visualize said chromosome 21 DNA.

10. The method of claim 9 wherein DNA of normal human chromosomes 1 through 20, 22, X, and Y is labeled with a second label that is distinguishable from the label used to visualize said chromosome 21 DNA.

11. The method of claim 1 wherein the labeled DNA comprises fragments complementary to a subregion of chromosome 21.

12. The method of claim 11 wherein DNA of normal human chromosomes 1 through 20, 22, X, and Y is labeled with a second label that is distinguishable from the label used to visualize said chromosome 21 DNA.

13. The method of claim 1 wherein the labeled unique sequence fragments comprise a fluorescent label.

14. The method of claim 1 wherein the labeled unique sequence fragments comprise an immunochemical label.

15. The method of claim 1 wherein the labeled unique sequence fragments comprise biotin.

16. The method of claim 1 wherein the heterogenous mixture of labeled unique sequence DNA fragments consists essentially of fragments complementary to human chromosome 21 DNA.