Method for the detection of chromosome structural abnormalities by in situ hybridization to fixed tissue
AssigneeOncor, Inc.
InventorsMary Ge Wang, Albert Louis George, Jr., Elizabeth Sophia Light
Life of this patent
Estimating the term…
Abstract
The present invention is directed to in situ hybridization methods using nucleic acid probes for single copy sequences for detecting chromosomal structural abnormalities in fixed tissue obtained from a patient suspected of having a chromosomal structural abnormality.
Claims42
A method of performing in situ hybridization with a probe which specifically hybridizes to a single copy sequence as a means of detecting a chromosome structural abnormality in a cell from a fixed tissue sample obtained from a patient suspected of having a chromosome structural abnormality wherein said method comprises the steps of:
- (a) obtaining a fixed tissue sample from said patient;
- (b) pretreating the fixed tissue sample obtained in step (a) with a bisulfite ion composition;
- (c) digesting the fixed tissue sample with proteinase;
- (d) performing in situ hybridization on cells obtained from the digested fixed tissue sample of step (c) with a probe which specifically hybridizes to a single copy sequence in a critical chromosomal region of interest wherein a signal pattern of hybridized probes is obtained;
- (e) comparing the signal pattern of the hybridized probe in step (d) to a predetermined signal pattern of the hybridized probe obtained when performing in situ hybridization on cells having a normal critical chromosome region of interest;
- and
- (f) detecting a chromosome structural abnormality in said critical chromosomal region of interest of the patient's cells, by detecting a difference between the signal pattern obtained in step (d) and the predetermined signal pattern.
A method of enhancing the signal generated using in situ hybridization with a probe for repetitive DNA in a cell from a fixed tissue sample wherein said method comprises the steps of:
- (a) obtaining a fixed tissue sample;
- (b) pretreating the fixed tissue sample obtained in step (a) with a bisulfite ion composition;
- (c) digesting the fixed tissue sample with an effective amount of proteinase;
- (d) performing in situ hybridization on cells obtained from the digested fixed tissue sample of step (c) with a probe for a repetitive DNA sequence in a critical chromosomal region of interest wherein a signal pattern of hybridized probes is obtained;
- (e) comparing the signal pattern of the hybridized probe in step (d) to a predetermined signal pattern of hybridized probe obtained when performing in situ hybridization on cells having a normal repetitive DNA region of interest;
- and (f) detecting a chromosome structural abnormality in said critical chromosomal region of interest, by detecting a difference between the signal pattern obtained in step (d) and the predetermined signal pattern.
Dependencies are inferred from the claim text.