Method for conducting multiparametric fluorescence in situ hybridization

• US 5,792,610 A
• Filed: 05/01/1996
• Issued: 08/11/1998
• Est. Priority Date: 05/01/1996
• Status: Expired due to Fees

• Current Assignee: Bio-Rad Laboratories Incorporated
• Original Assignee: Bio-Rad Laboratories Incorporated
• Inventors: Heffelfinger, David M., Witney, Franklin R.
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): Atzel, Amy
• Application Number:
  US08/640,658
• Time in Patent Office: 832 Days
• Field of Search: 435/6, 356/317, 356/318, 356/345, 356/346, 356/359, 356/363
• US Class Current: 435/6.13
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6841 In situ hybridisation

Abstract

The invention relates to a method of identifying the individual autosomal and sex chromosomes of a human karyotype through the use of a set of combinatorially labeled oligonucleotide probes each member thereof: (i) having a predetermined label distinguishable from the label of any other member of said set, and (ii) being capable of specifically hybridizing with one predetermined autosomal or sex chromosome of a human karyotype.

Claims

1. A method of simultaneously identifying and distinguishing the individual autosomal and sex chromosomes of a human karyotype which comprises the steps:

(a) contacting a preparation of said chromosomes, in single-stranded form, under conditions sufficient to permit nucleic acid hybridization to occur with a set of combinatorially labeled oligonucleotide probes, each member thereof;

(i) having a predetermined fluorescent label distinguishable from the label of any other member of said set, and (ii) being capable of specifically hybridizing with one predetermined autosomal or sex chromosome of a human karyotype;

said set having sufficient members to be capable of specifically hybridizing each autosomal and sex chromosome of said human karyotype;

wherein said contacting thereby results in each autosomal or sex chromosome of said preparation being hybridized to at least one member of said set of probes;

(b) for each chromosome of said preparation hybridized to at least one member of said set of probes, employing an interferometer to detect and identify the predetermined label of the hybridized probe member(s) to correlate the identity of the label with the identity of the autosomal and sex chromosomes of said human karyotype, thereby identifying the chromosome hybridized to said probe member(s); and

(c) repeating step (b) until each autosomal and sex chromosome of said human karyotype has been identified in said preparation.

2. The method of claim 1, wherein said interferometer is a common path interferometer.

3. The method of claim 2, wherein said common path interferometer is a Sagnac interferometer.

4. The method of claim 1, wherein, in step (b), an interferogram is produced from said interferometer, and said interferogram is Fourier transformed to recover a spectral signature of the predetermined label of said hybridized probe member.

5. The method of claim 4, wherein the spectral signature of the predetermined label of said hybridized probe member in step (b) is recovered by comparing said interferogram to a library or lookup table of previously determined interferograms.

6. The method of claim 2, wherein in step (b), an interferogram is produced from said interferometer, and said interferogram is Fourier transformed to recover a spectral signature of the predetermined label of said hybridized probe member.

7. The method of claim 6, wherein the spectral signature of the predetermined label of said hybridized probe member in step (b) is recovered by comparing said interferogram to a library or lookup table of previously determined interferograms.

8. The method of claim 3, wherein, in step (b), an interferogram is produced from said interferometer, and said interferogram is Fourier transformed to recover the spectral signature of the predetermined label of said hybridized probe member.

9. The method of claim 8, wherein the spectral signature of the predetermined label of said hybridized probe member in step (b) is recovered by comparing said interferogram to a library or lookup table of previously determined interferograms.