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1. WO2001079534 - PROCEDE DE DETECTION DE CELLULES NEOPLASIQUES ET NON NEOPLASIQUES

Note: Texte fondé sur des processus automatiques de reconnaissance optique de caractères. Seule la version PDF a une valeur juridique

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CLAIMS:

1 . A method for monitoring a lymphoid neoplastic condition in a mammal, said method comprising contacting the nucleic acid molecules contained in a sample derived from said mammal with a nucleic acid reference molecule or derivative or analogue thereof which is complementary to a marker nucleic acid molecule or analogue thereof, which marker molecule is characteristic of the neoplastic cells, for a time and under conditions sufficient to facilitate the interaction of said reference molecule with said marker molecule, enriching for said marker molecule by reducing the concentration of un-hybridised nucleic acid molecules and hybridisation-mismatched nucleic acid molecules and qualitatively and/or quantitatively detecting said enriched marker nucleic acid molecules.

2. The method according to claim 1 wherein said reference nucleic acid molecule is a driver nucleic acid molecule.

3. The method according to claim 1 or 2 wherein the step of enriching for the marker molecule by reducing concentrations of un-hybridised nucleic acid molecules and hybridisation-mismatched nucleic acid molecules further includes the step of reducing the concentration of reference nucleic acid molecules.

4. The method according to claim 3 wherein the enrichment step is performed by removing non-marker nucleic acid molecules which have not hybridised to a tagged driver nucleic acid molecule, enzymatically cleaving any mismatched hybridisation nucleic acid molecules, and removing driver molecules.

5. The method according to claim 3 wherein the enrichment step is performed by separating reference marker homoduplexes from hybridisation and mismatched heteroduplexes based on differential migration of the duplexes through a gel or a size exclusion or affinity or other matrix.

6. The method according to claim 5 wherein said matrix is contained in a column or capillary.

7. The method according to claim 4 wherein said driver molecules are removed by the use of UNG.

8. The method according to any one of claims 1-7 wherein said marker is a rearranged TCR or immunoglobulin variable region nucleic acid molecule or derivative or analogue thereof.

9. The method according to claim 8 wherein said variable region nucleic acid molecule is the genomic form of the rearranged variable region gene segment.

10. The method according to claim 9 wherein said lymphoid neoplastic condition is a lymphoid malignant condition.

11. The method according to claim 10 wherein said monitoring is monitoring of the minimal residual disease condition.

12. A method for detecting and/or quantifying a clonal population of cells in a biological sample said cells being characterised by a marker nucleic acid molecule, which marker nucleic acid molecule is electrophoretically co-migratable within said population of cells, said method comprising electrophoretically separating the nucleic acid molecules contained in said sample, wherein said separation is based on nucleic acid length and sequence, and detecting said separated nucleic acid molecules.

13. The method according to claim 12 wherein said clonal population of cells is a population of neoplastic cells.

14. The method according to claim 13 wherein said neoplastic cells are malignant.

15. The method according to claim 13 or 14 wherein said neoplastic cells are neoplastic lymphoid cells.

16. The method according to claim 15 wherein said marker is a rearranged TCR or immunoglobulin variable region nucleic acid molecule or derivative or analogue thereof.

17. The method according to claim 16 wherein said variable region nucleic acid molecule is the genomic form of the rearranged variable region gene segment.

18. The method according to any one of claims 12-17 wherein said separation is two dimensional denaturing gradient gel electrophoresis.

19. The method according to anyone of claims 12-18 wherein the subject neoplastic condition is the clonal evolution of a neoplastic cell.

20. A method for detecting and/or quantifying multiple non-neoplastic lymphoid cells in a biological sample, said non-neoplastic cells being characterised by a marker nucleic acid molecule, which marker nucleic acid molecule is electrophoretically co-migratable within said population of cells, said method comprising electrophoretically separating the nucleic acid molecules contained in said sample, wherein said separation is based on nucleic acid length and sequence, and detecting said separated nucleic acid molecules.

21. The method according to claim 20 wherein said separation is two dimensional denaturing gradient gel electrophoresis.

22. The method according to any one of claims 1-21 substantially as hereinbefore described with reference to the Figures and/or Examples.