Methods for detection of genetic disorders
OwnerRavinder Dhallan, Xin Guoinventors
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Abstract
The invention provides a method useful for detection of genetic disorders. The method comprises determining the sequence of alleles of a locus of interest, and quantitating a ratio for the alleles at the locus of interest, wherein the ratio indicates the presence or absence of a chromosomal abnormality. The present invention also provides a non-invasive method for the detection of chromosomal abnormalities in a fetus. The invention is especially useful as a non-invasive method for determining the sequence of fetal DNA. The invention further provides methods of isolation of free DNA from a sample.
Claims23
A method for determining the sequence of alleles at a locus of interest from template DNA in a sample, said method comprising:
- obtaining a non-cellular fraction of said sample, concentrating DNA in said non-cellular fraction, and determining the sequence of alleles at the locus of interest in said concentrated sample.
A method for detecting a chromosomal abnormality in a sample, said method comprising:
- (a) obtaining a non-cellular fraction of said sample,
- (b) concentrating template DNA within said non-cellular fraction,
- (c) quantitating the relative amount of alleles at a heterozygous locus of interest, wherein said heterozygous locus of interest was identified by determining the sequence of alleles at a locus of interest from template DNA, wherein said relative amount is expressed as a ratio, and wherein said ratio indicates the presence or absence of a chromosomal abnormality.
A method for determining the sequence of alleles at a locus of interest from template DNA in a sample to which a cell lysis inhibitor has been added comprising:
- (a) obtain a non-cellular fraction of said sample;
- (b) concentrating the template DNA in said non-cellular fraction;
- (c) amplifying alleles of a locus of interest on said concentrated template DNA using a first and a second primer, wherein the second primer contains a recognition site for a restriction enzyme such that digestion with the restriction enzyme generates a 5′ overhang containing the locus of interest;
- (d) digesting the amplified DNA with the restriction enzyme that recognizes the recognition site on the second primer;
- (e) incorporating a nucleotide into the digested DNA of (b) by using the 5′ overhang containing the locus of interest as a template;
- and
- (f) determining the sequence of the alleles of the locus of interest by determining the sequence of the DNA of (c).
Dependencies are inferred from the claim text.