US 11,879,157 B2
Target-enriched multiplexed parallel analysis for assessment of risk for genetic conditions
George Koumbaris, Lithrodontas (CY); Marios Ioannides, Nicosia (CY); Kyriakos Tsangaras, Limassol (CY); Charalambos Loizides, Nicosia (CY); Michalis Nicolaou, Nicosia (CY); Skevi Kyriakou, Larnaca (CY); Philippos Patsalis, Nicosia (CY); and Elena Kypri, Nico (CY)
Assigned to NIPD GENETICS PUBLIC COMPANY LIMITED, Nicosia (CY)
Appl. No. 16/625,440
Filed by NIPD GENETICS PUBLIC COMPANY LIMITED, Nicosia (CY)
PCT Filed Jul. 6, 2018, PCT No. PCT/EP2018/068431
§ 371(c)(1), (2) Date Dec. 20, 2019,
PCT Pub. No. WO2019/008167, PCT Pub. Date Jan. 10, 2019.
Claims priority of provisional application 62/529,785, filed on Jul. 7, 2017.
Prior Publication US 2021/0147937 A1, May 20, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/6883 (2018.01); C12Q 1/6874 (2018.01)
CPC C12Q 1/6883 (2013.01) [C12Q 1/6874 (2013.01); C12Q 2600/112 (2013.01); C12Q 2600/118 (2013.01)] 18 Claims
 
1. A method of determining status of a genetic condition or a fetal risk of inheriting a genetic condition, the method comprising:
(a) preparing a sequencing library from a sample comprising maternal and fetal DNA;
(b) hybridizing the sequencing library to a pool of single-stranded TArget Capture Sequences (TACS), wherein the pool of TACS comprises a plurality of sequences that bind to genomic sequences of interest including variant allele loci of interest associated with different genetic conditions;
(c) isolating members of the sequencing library that bind to the pool of TACS to obtain an enriched library;
(d) amplifying and sequencing the enriched library;
(e) performing statistical analysis on the enriched library sequences to thereby determine maternal carrier status at the loci of interest associated with different genetic conditions;
(f) selecting, after step (e), the sample comprising maternal and fetal DNA carrying a variant allele associated with an inheritable genetic condition;
(g) obtaining a paternal DNA sample and performing steps (a)-(e) on the paternal DNA sample to determine paternal carrier status;
(h) selecting, after step (g), the paternal DNA sample carrying a variant allele associated with an inheritable genetic condition in common with the sample comprising maternal and fetal DNA carrying the variant allele associated with the inheritable genetic condition; and
(i) determining fetal risk of inheriting a genetic condition based on maternal carrier status selected in step (f) and paternal carrier status selected in step (h).