Methods and nucleic acids for analyses of cellular proliferative disorders
View Patent ↗Aspects of the invention provide methods, nucleic acids and kits for detecting, or for detecting and distinguishing between or among liver cell proliferative disorders or for detecting, or for detecting and distinguishing between or among colorectal cell proliferative disorders. Particular aspects disclose and provide genomic sequences the methylation patterns of which have substantial utility for the improved detection of and differentiation between said class of disorders, thereby enabling the improved diagnosis and treatment of patients.
1. An isolated nucleic acid molecule selected from the group consisting of SEQ ID NOs: 30, 31, 42, and 43, wherein the nucleic acid molecule is not identical or complementary to all or a portion of SEQ ID NO: 24 or other naturally occurring DNA.
2. An isolated nucleic acid molecule, comprising at least 16 contiguous nucleotides of a DNA sequence selected from the group consisting of SEQ ID NOS:30, 31, 42, and 43, and sequences fully complementary to the isolated nucleic acid, wherein the sequence:
(i) is chemically linked to an entity selected from the group consisting of chromophores, fluorophores, lipids, cholic acid, thioethers, aliphatic chains phospholipids, polyamines and polyethylene glycol, or
(ii) is bound to a solid phase support.
3. An isolated nucleic acid molecule, comprising at least 50 contiguous nucleotides of a DNA sequence selected from the group consisting of SEQ ID NOS:30, 31, 42, and 43 and sequences fully complementary thereto.
4. The nucleic acid of claim 1 wherein the contiguous base sequence comprises at least one CpG, TpG or CpA dinucleotide sequence.
5. The nucleic acid of claim 2 wherein the contiguous base sequence comprises at least one CpG, TpG or CpA dinucleotide sequence.
6. The nucleic acid of claim 3 wherein the contiguous base sequence comprises at least one CpG, TpG or CpA dinucleotide sequence.
7. A kit suitable for detecting and/or classifying cellular proliferative disorders in a subject, comprising (a) a bisulfite reagent; (b) a container suitable for containing the bisulfite reagent and a biological sample of the subject; (c) at least one oligonucleotide whose sequence is identical, fully complementary, or hybridizes when incubated at 68° C. in 5×SSC/5×Denhardt's solution and 1.0% SDS and washed at room temperature in 0.2×SSC/0.1% SDS to an 18 base long segment of a sequence selected from SEQ ID NOS:42 and 43, wherein the sequence comprises at least one TpA or CpA dinucleotide.
8. An isolated amplificate nucleic acid comprising at least 50 nucleotides of a nucleic acid sequence selected from SEQ ID NOs: 31, 42 and 43.