Molecular diagnostics for galactosemia
AssigneeEmory University
InventorsLouis J. Elsas, II, Kasinathan Muralidharan
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Abstract
A process for detecting mutations in the gene responsible for galactosemia, galactose-1-phosphate uridyl transferase (GALT), is described. In one embodiment, the process can be used to detect over 85% of the mutations known to cause galactosemia in the United States population by using six different oligonucleotide probes, which span single-nucleotide Missense or nonsense mutations in the GALT gene. Hybridization conditions which can distinguish a single nucleotide mismatch are used to detect both the presence and zygosity of mutations in the GALT gene to aid in genetic counseling. A kit for use in detecting mutations in the GALT gene is also disclosed.
Claims11
A process for detecting a mutation in the GALT gene that causes galactosemia, comprising
- (a) amplifying the GALT gene in a patient's genomic DNA,
- (b) hybridizing the amplified DNA with a probe at least 10 nucleotides in length that comprises sequence complementary to GCCTGTAAG, and
- (c) detecting the presence or absence of a hybrid, wherein the presence of the hybrid indicates the presence of a galactosemia-causing mutation.
A process for detecting mutations in the GALT gene that cause galactosemia, comprising
- (a) amplifying the GALT gene in a patient's genomic DNA,
- (b) hybridizing the amplified DNA, serially or in parallel, with each member of a set of detectable probes, each probe of the set being at least 10 nucleotides in length, the set comprising probes having sequence complementary to the following sequences:
- TGGTTGGAT, TGCCGGGTA, TTCCCGCCA, CCAATTATG, GCCTGTAAG, and CTGGGACCA, under hybridization conditions that allow only exactly complementary strands to remain as hybrids, and
- (c) detecting the presence or absence of the hybrids, wherein the presence of one or more of the hybrids indicates the presence of one or more galactosemia-causing mutations.
A process for detecting mutations in the GALT gene that cause galactosemia, comprising
- (a) amplifying the GALT gene in a patient's genomic DNA,
- (b) hybridizing the amplified DNA, serially or in parallel, with each member of a set of detectable probes, each probe of the set being at least 10 nucleotides in length, the set comprising probes having sequence complementary to the following sequences:
- TGGTTGGAT, TGCCGGGTA, TTCCCGCCA, CCAATTATG, GCCTGTAAG, and CTGGGACCA, under hybridization conditions that allow both exact and mismatched hybrids to form, and,
- (c) determining the melting temperature of the hybrids formed in (b), wherein the value of the melting temperature indicates whether no, one or more galoctosemia-causing mutations are present.
Dependencies are inferred from the claim text.