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Method of analyzing chromosomal inversions

  • US 9,562,259 B2
  • Filed: 03/12/2012
  • Issued: 02/07/2017
  • Est. Priority Date: 03/14/2011
  • Status: Active Grant
First Claim
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1. A chromogenic in situ hybridization method for determining whether a sample comprises a chromosomal rearrangement, the chromosomal rearrangement occurring as a result of a breakpoint within a gene, comprising:

  • contacting the sample witha first nucleic acid probe comprising a first sequence configured to hybridize to a first chromosomal DNA target located 5′

    to the breakpoint,a second nucleic acid probe comprising a second sequence configured to hybridize to a second chromosomal DNA target located 3′

    to the breakpoint, anda third nucleic acid probe comprising a third sequence having a 5′

    portion and a 3′

    portion, the 5′

    portion configured to hybridize to a portion of a third chromosomal DNA target that is 5′ and

    adjacent to the breakpoint, and the 3′

    portion configured to hybridize to a portion of the third chromosomal DNA target that is 3′ and

    adjacent to the breakpoint, such that in an absence of a rearrangement the third nucleic acid probe hybridizes to a region of the third chromosomal DNA target spanning the breakpoint;

    establishing conditions suitable for the first, second, and third probes to hybridize to the respective chromosomal DNA targets in the sample;

    contacting the sample with first, second, and third detection reagents, the first detection reagent comprising components to label the first chromosomal DNA target with a first chromogen, the second detection reagent comprising components to label the second chromosomal DNA target with a second chromogen, and the third detection reagent comprising components to label the third chromosomal DNA target with a third chromogen, where each of the first, second, and third chromogens provide different detectable signals;

    detecting a colocalization of a third signal from the third labeled chromosomal DNA target with a first signal from the first labeled chromosomal DNA target;

    detecting a colocalization of the third signal from the third labeled chromosomal DNA target with a second signal from the second labeled chromosomal DNA target; and

    identifying the chromosomal rearrangement based on the detected colocalizations.

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