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 man
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), wherein the morphological stain used in step (a) does not significantly interfere with detection of the labeled nucleic acid probe used in step (e). 2. The method of claim 1 wherein the label is a label selected from the group consisting of a fluorescent dye, a colloidal gold particle, a radioisotope label, and an enzyme substrate label. 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 selected from the group consisting of HER2/neu gene and the centromere of chromosome 17. 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 label is a radioisotope label selected from the group consisting of 35S, 14C, 125I, 3H, and 131I. 9. The method of claim 1, wherein the label is 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). 10. The method of claim 1, wherein the label is a fluorescent dye selected from the group consisting of Texas Red, fluorescein, phycocrytherin, rhodamine, phycocyanin, dansyl, umbelliferone, SPECTRUM GREEN®, and SPECTRUM ORANGE®. 11. 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), wherein autofluorescence of the morphological stain used in step (a) does not significantly interfere with detection of the labeled nucleic acid probe used in step (e). 12. The method of claim 11 wherein the label is a fluorescent dye. 13. The method of claim 11, 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. 14. The method of claim 11, wherein the target nucleic acid is selected from HER2/neu gene and the centromere of chromosome 17. 15. The method of claim 11, wherein the target protein is a HER2 protein. 16. The method of claim 11, wherein the tissue sample is selected from the group consisting of breast, prostate, ovary, colon, lung, endometrium, stomach, salivary gland and pancreas tissue samples. 17. The method of claim 11, wherein the section of tissue sample is obtained from fixed, paraffin-embedded tissue sample. 18. 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), wherein the morphological stain used in step (a) does not significantly autofluoresce at the same wavelength as a fluorescent label of the fluorescently labeled nucleic acid probe used in step (e). 19. The method of claim 18 wherein the label is a fluorescent dye. 20. The method of claim 18, 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. 21. The method of claim 18, wherein the target nucleic acid is selected from HER2/neu gene and the centromere of chromosome 17. 22. The method of claim 18, wherein the target protein is a HER2 protein. 23. The method of claim 18, wherein the tissue sample is selected from the group consisting of breast, prostate, ovary, colon, lung, endometrium, stomach, salivary gland and pancreas tissue samples. 24. The method of claim 18, wherein the section of tissue sample is obtained from fixed, paraffin-embedded tissue sample.
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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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