Methods for fetal DNA detection and allele quantitation
OwnerGregory M. Landes, Lesley Michalowsky +2inventors
InventorsGregory M. Landes, Lesley Michalowsky, Glenn Miller, William Weber
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Abstract
The present invention provides non-invasive methods to distinguish fetal DNA from maternal DNA and thereby detect fetal aneuploidies and alleles. The methods require isolation of fetal DNA from maternal serum and treatment with a reagent that creates primary sequence differences between maternal and fetal DNA that exhibit differential methylation. Various methods including quantitative PCR is used to identify detect fetal aneuploidies and alleles. In one embodiment, the method is useful to identify imprinting genes in subjects, including adults.
Claims4
A method of detecting fetal aneuploidies comprising
- a) treating DNA isolated from maternal serum with a reagent that differentially modifies methylated and non-methylated DNA;
- b) performing quantitative PCR with a first primer pair on a potentially aneuploid chromosome;
- c) performing a control quantitative PCR with a second primer pair on a non-aneuploid chromosome;
- and
- d) determining the ratio of the quantity of the two PCR products, thereby detecting fetal aneuploidies.
A method for detecting fetal chromosome aneuploidies comprising:
- a) treating DNA isolated from maternal serum with bisulfite;
- b) performing quantitative PCR on the sample with a primer pair homologous to a test chromosome sequence that is differentially methylated in maternal DNA and in fetal DNA, where the primer pair only primes bisulfite treated unmethylated DNA;
- c) performing a control quantitative PCR with a primer pair homologous to a control chromosome sequence that is differentially methylated in maternal DNA and in fetal DNA, where the primer pair only primes bisulfite treated unmethylated DNA;
- and
- d) determining the ratio of the quantity of PCR product produced for the test chromosome compared with the control chromosome.
A method for detecting alleles of a gene of interest in fetal DNA comprising:
- a) treating DNA isolated from maternal serum with bisulfite;
- b) performing PCR with a primer pair that amplifies the gene of interest of step b);
- and
- c) analyzing the resulting PCR product to identify the allele of the gene of interest.
A method for detecting imprinted genes in a subject comprising:
- a) treating DNA isolated from a subject with bisulfite;
- b) performing PCR with a primer pair for a polymorphic region that only amplified bisulfite treated unmethylated DNA;
- and
- c) analyzing the PCR product to identify the polymorphism thereby detecting imprinted genes in a subject.
Dependencies are inferred from the claim text.