US 11,674,176 B2
Fetal aneuploidy detection by sequencing
Roland Stoughton, The Sea Ranch, CA (US); Ravi Kapur, Sharon, MA (US); Barb Ariel Cohen, Watertown, MA (US); and Mehmet Toner, Charlestown, MA (US)
Assigned to Verinata Health, Inc, Redwood City, CA (US); The General Hospital Corporation, Boston, MA (US); and GPB Scientific, LLC, Richmond, VA (US)
Filed by Verinata Health, Inc., Redwood City, CA (US); The General Hospital Corporation, Boston, MA (US); and GPB Scientific, LLC, Richmond, VA (US)
Filed on Jun. 30, 2020, as Appl. No. 16/917,185.
Application 16/917,185 is a continuation of application No. 13/794,503, filed on Mar. 11, 2013, granted, now 10,704,090.
Application 13/794,503 is a continuation of application No. 12/751,940, filed on Mar. 31, 2010, abandoned.
Application 12/751,940 is a continuation of application No. 11/763,133, filed on Jun. 14, 2007, abandoned.
Claims priority of provisional application 60/820,778, filed on Jul. 28, 2006.
Claims priority of provisional application 60/804,816, filed on Jun. 14, 2006.
Prior Publication US 2021/0087624 A1, Mar. 25, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/6874 (2018.01); C12Q 1/6883 (2018.01)
CPC C12Q 1/6874 (2013.01) [C12Q 1/6883 (2013.01); C12Q 2600/156 (2013.01)] 11 Claims
 
1. A method for determining a ratio of signal intensities of amplification products for a maternal allele in a first genomic region on a first chromosome suspected of being aneuploid and for a maternal allele in a second genomic region on a second chromosome that is not aneuploid or not suspected of being aneuploid in a mixed sample comprising a mixture of fetal and maternal cells, the method comprising:
a. enriching the mixed sample for fetal cells to produce an enriched sample comprising fetal and maternal cells having an increased concentration of fetal cells relative to maternal cells before enrichment, and lysing cells in the enriched sample to obtain genomic DNA (gDNA);
b. hybridizing probes to each of a plurality of loci in the gDNA on at least two different chromosomes to produce hybridized probes;
c. ligating the hybridized probes at each locus to each other to create an amplification template specific to each locus;
d. amplifying the amplification template specific to each locus to create amplification products;
e. hybridizing the amplification products to probes on a microarray; and
f. detecting, among the amplification products hybridized to probes on the microarray, a first signal intensity of the amplification products for a maternal allele in a first genomic region on a first chromosome suspected of being aneuploid and a second signal intensity of the amplification products for a maternal allele in a second genomic region on a second chromosome that is not aneuploid or not suspected of being aneuploid, and determining a ratio of the first signal intensity to the second signal intensity.