Methods for the diagnosis of fetal abnormalities
First Claim
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1. A method for determining the presence or absence of a fetal aneuploidy using a maternal blood sample comprising a mixture of fetal and maternal genomic nucleic acids, the method comprising:
- a. obtaining a test sample comprising a mixture of fetal and maternal genomic nucleic acids from a maternal blood sample;
b. selectively amplifying a plurality of target nucleic acids from the test sample and a plurality of target nucleic acids from a reference sample using primers labeled with a unique label for each target region, thereby generating amplification products, wherein the target nucleic acids are selected from one or more chromosomes to be tested for aneuploidy, and wherein the reference sample is diploid for the one or more chromosomes to be tested for aneuploidy;
c. amplifying the amplification products using universal primers;
d. detecting the amplified target nucleic acids from the test sample and from the reference sample;
e. quantifying the detected amplified target nucleic acids from the test sample and from the reference sample;
f. analyzing the test sample for the presence or absence of a fetal aneuploidy by comparing the quantity of the detected amplified target nucleic acids obtained from the test sample for the one or more chromosomes tested for aneuploidy to the quantity of the detected amplified target nucleic acids obtained from the reference sample for the one or more chromosomes tested for aneuploidy; and
g. generating a report on the presence or absence of aneuploidy in the maternal blood sample.
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Abstract
The present invention relates to methods for detecting, enriching, and analyzing rare cells that are present in the blood, e.g. fetal cells. The invention further features methods of analyzing rare cell(s) to determine the presence of an abnormality, disease or condition in a subject, e.g. a fetus by analyzing a cellular sample from the subject.
63 Citations
26 Claims
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1. A method for determining the presence or absence of a fetal aneuploidy using a maternal blood sample comprising a mixture of fetal and maternal genomic nucleic acids, the method comprising:
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a. obtaining a test sample comprising a mixture of fetal and maternal genomic nucleic acids from a maternal blood sample; b. selectively amplifying a plurality of target nucleic acids from the test sample and a plurality of target nucleic acids from a reference sample using primers labeled with a unique label for each target region, thereby generating amplification products, wherein the target nucleic acids are selected from one or more chromosomes to be tested for aneuploidy, and wherein the reference sample is diploid for the one or more chromosomes to be tested for aneuploidy; c. amplifying the amplification products using universal primers; d. detecting the amplified target nucleic acids from the test sample and from the reference sample; e. quantifying the detected amplified target nucleic acids from the test sample and from the reference sample; f. analyzing the test sample for the presence or absence of a fetal aneuploidy by comparing the quantity of the detected amplified target nucleic acids obtained from the test sample for the one or more chromosomes tested for aneuploidy to the quantity of the detected amplified target nucleic acids obtained from the reference sample for the one or more chromosomes tested for aneuploidy; and g. generating a report on the presence or absence of aneuploidy in the maternal blood sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
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16. A method of determining the presence or absence of a fetal aneuploidy using a maternal blood sample comprising a mixture of fetal and maternal genomic nucleic acids, the method comprising:
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a. obtaining a mixture of fetal and maternal genomic nucleic acids from a maternal blood sample; b. hybridizing genomic nucleic acids with one set of oligonucleotides per target nucleic acid, wherein each set comprises two or more oligonucleotides that each hybridize to different regions within the target nucleic acid and wherein each oligonucleotide comprises a universal polymerase chain reaction (PCR) primer site to produce hybridization products; c. processing the hybridization products to provide a template for amplification; d. amplifying the hybridization products using primers with a unique label for each target region, thereby generating amplification products; e. amplifying the amplification products using universal primers; f. hybridizing the single-stranded, labeled nucleic acids to complementary nucleic acids on an array or bead; g. detecting labels on the array or beads and analyzing the labels to determine genotypes of the nucleic acids; and h. generating a report on the presence or absence of aneuploidy in the maternal blood sample based on the genotype analysis. - View Dependent Claims (17, 18, 19, 20, 21, 22, 23, 24, 25, 26)
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Specification