Rapid aneuploidy detection
AssigneeThe Johns Hopkins University
InventorsBert Vogelstein, Kenneth W. Kinzler, Nickolas Papadopoulos, Isaac A. Kinde
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Abstract
Massively parallel sequencing of cell-free, maternal plasma DNA was recently demonstrated to be a safe and effective screening method for fetal chromosomal aneuploidies. Here, we report an improved sequencing method achieving significantly increased throughput and decreased cost by replacing laborious sequencing library preparation steps with PCR employing a single primer pair. Using this approach, samples containing as little as 4% trisomy 21 DNA could be readily distinguished from euploid samples.
Claims12
(canceled)
A method of detecting the presence or absence of aneuploidy, comprising:
- a) contacting a sample comprising nucleic acid from a human with a single primer pair, wherein said single primer pair is designed to anneal to a plurality of repeat genomic regions present on a reference set of all twenty-two autosome human non-aneuploid chromosomes that would generate, during amplification, an expected distribution, b) amplifying, using polymerase chain reaction, the nucleic acid using the single primer pair, whereby a plurality of amplicons are produced, wherein said amplicons are distinct from one another;
- c) sequencing at least 5 nucleotides of each of the amplicons in the plurality of amplicons;
- d) aligning in silico the sequenced amplicons to said reference set of all twenty-two autosome human non-aneuploid chromosomes in order to produce a query distribution;
- and
- e) identifying the presence or absence of aneuploidy based upon comparing said query distribution to said expected distribution.
Dependencies are inferred from the claim text.