This invention relates to methods of analyzing a tissue sample from a subject. In particular, the invention combines morphological staining and/or immunohistochemistry (IHC) with fluorescence in situ hybridization (FISH) within the same section of a tissue sample. The analysis can be automated or ma
This invention relates to methods of analyzing a tissue sample from a subject. In particular, the invention combines morphological staining and/or immunohistochemistry (IHC) with fluorescence in situ hybridization (FISH) within the same section of a tissue sample. The analysis can be automated or manual. The invention also relates to kits for use in the above methods.
대표청구항▼
What is claimed is: 1. A method for detecting the presence of a cellular target protein and a cellular target nucleic acid in a locus of interest in a tissue sample containing cells of interest, wherein said cells of interest may comprise normal cells or may comprise abnormal cells, or both, and sa
What is claimed is: 1. A method for detecting the presence of a cellular target protein and a cellular target nucleic acid in a locus of interest in a tissue sample containing cells of interest, wherein said cells of interest may comprise normal cells or may comprise abnormal cells, or both, and said locus of interest comprises a region containing cells of interest, said method comprising the steps of: (a) applying a morphological stain to a tissue sample effective to provide a stained tissue sample; (b) identifying a locus of interest in said stained tissue sample; (c) contacting a section of the tissue sample with an antibody which specifically binds to the target protein, wherein said section includes said locus of interest; (d) determining binding of the antibody to the section of tissue sample; (e) hybridizing a labeled nucleic acid probe to the target nucleic acid in the same section of tissue sample which includes the locus of interest; (f) detecting the presence of the labeled nucleic acid probe in the section of tissue sample; and (g) comparing the results of step (d) with the results of step (f). 2. The method of claim 1 wherein the label is a fluorescent dye. 3. The method of claim 1, wherein the nucleic acid probe is constructed to hybridize to the target nucleic acid sequence indicating a genetic abnormality selected from the group consisting of amplification, addition, substitution, translocation and deletion. 4. The method of claim 1, wherein the target nucleic acid is a HER2/neu gene. 5. The method of claim 1, wherein the target protein is a HER2 protein. 6. The method of claim 1, wherein the tissue sample is selected from the group consisting of breast, prostate, ovary, colon, lung, endometrium, stomach, salivary gland and pancreas tissue samples. 7. The method of claim 1, wherein the section of tissue sample is obtained from fixed, paraffin-embedded tissue sample. 8. The method of claim 1, wherein the section of tissue sample is not destained prior to step (c). 9. The method of claim 1, wherein the morphological stain used in step (a) does not significantly autofluoresce at the same wavelength as a fluorescent label of the first nucleic acid probe. 10. The method of claim 1, wherein the morphological stain comprises xanthine dye and thiazine dye. 11. The method of claim 1, wherein the morphological stain comprises methylene blue, a xanthine dye, or a thiazine dye. 12. The method of claim 1, wherein the morphological stain is HEMA 3��. 13. The method of claim 1, further comprising hybridizing a second labeled nucleic acid probe to a nucleic acid sequence in the section of tissue sample, wherein the second nucleic acid probe comprises a label distinguishable from a label of the first nucleic acid probe. 14. The method of claim 13, wherein the second nucleic acid probe determines chromosome copy number. 15. The method of claim 13, wherein said first and said second labeled nucleic acid probe are fluorescently labeled nucleic acid probes, and wherein said second labeled nucleic acid probe comprises a fluorescent label distinguishable from a fluorescent label of the first nucleic acid probe. 16. The method of claim 1, wherein the nucleic acid probe is labeled with a radioisotope label selected from the group consisting of 35S, 14C, 125l 3H, and 131l. 17. The method of claim 1, wherein the nucleic acid probe is labeled with colloidal gold particles. 18. The method of claim 1, wherein the nucleic acid probe is labeled with an enzyme substrate label selected from the group of enzyme substrate labels consisting of horseradish peroxidase (HRPO), alkaline phosphatase (AP), and β-D-galactosidase (β-D-Gal). 19. The method of claim 1, wherein the target nucleic acid sequence is selected from the group consisting of HER2/neu gene and the centromere of chromosome 17. 20. The method of claim 1, wherein the nucleic acid probe is labeled with a fluorescent label selected from the group consisting of Texas Red, fluorescein, phycocrytherin, rhodamine, phycocyanin, dansyl, umbelliferone, SPECTRUM GREEN��, and SPECTRUM ORANGE��.
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이 특허에 인용된 특허 (6)
Slamon Dennis J. (Woodland Hills CA) McGuire William L. (San Antonio TX), Determination of status in neoplastic disease.
Cheever Martin A. ; Disis Mary L., Immune reactivity to HER-2/neu protein for diagnosis and treatment of malignancies in which the HER-2/neu oncogene is a.
Cheever Martin A. ; Disis Mary L., Immune reactivity to HER-2/neu protein for diagnosis and treatment of malignancies in which the HER-2/neu oncogene is a.
Tryggvason Karl (Fyysikontic 8 ; FIN-90570 Oulu FIX) Kallunki Pekka (8722 La Jolla Dr. ; Unit 99 La Jolla CA 92037) Pyke Charles (Solbakken 4 ; DK-3400 Hilleroo DKX), Laminin chains: diagnostic uses.
Ness Jeffrey Van ; Tabone John C. ; Howbert J. Jeffry ; Mulligan John T., Methods and compositions for enhancing sensitivity in the analysis of biological-based assays.
Slamon Dennis J. ; Press Michael F., Quantitative measurement of tissue protein identified by immunohistochemistry and standardized protein determination.
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