INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) - (19) World Intellectual Property Organization - - 1111111111111111111111111111111111111111111111111111111111111111111111111110111111 ' International Bureau (10) International Publication Number (43) International Publication Date ......0\'\' WO 2018/072705 Al 26 April 2018 (26.04.2018) WIP0 I PCT (51) International Patent Classification: Yuen Road, Shatin, New Territories, Hong Kong 999077 C12Q 1/68 (2006.01) (CN). (21) International Application Number: (74) Agent: INSIGHT INTELLECTUAL PROPERTY PCT/CN2017/106663 LIMITED; 19 A, Tower A, InDo Building, No. 48A (22) International Filing Date: Zhichun Road, Haidian District, Beijing 100098 (CN). 18 October 2017 (18.10.2017) (81) Designated States (unless otherwise indicated, for every (25) Filing Language: English kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, (26) Publication Language: English CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, (30) Priority Data: DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, 62/410,108 19 October 2016 (19.10.2016) US HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, (71) Applicant: THE CHINESE UNIVERSITY OF HONG MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, KONG [CN/CN]; c/o Office of Research and Knowledge OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, Transfer Services, Room 301, Pi Ch'iu Building, Shatin, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, New Territories, Hong Kong 999077 (CN). TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (72) Inventors: LO, Yuk-Ming Dennis; 4th Floor, 7 King Tak (84) Designated States (unless otherwise indicated, for every Street, Homantin, Kowloon, Hong Kong 999077 (CN). kind of regional protection available): ARIPO (BW, GH, _ CHIU, Rossa Wai Kwun; House 31, Double Haven, 52 GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, Ma Lok Path, Shatin, New Territories, Hong Kong 999077 UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, (CN). CHAN, Kwan Chee; Flat C, 15th Floor, Block 5, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, Peak One, Mei Tin Road, Shatin, New Territories, Hong EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, = Kong 999077 (CN). JIANG, Peiyong; Flat 7 on 1st Floor of MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, = _ Block B of Kwong Lam Court, Nos. 62-66 (No.62) Siu Lek = (54) Title: GESTATIONAL AGE ASSESSMENT BY METHYLATION AND SIZE PROFILING OF MATERNAL PLASMA DNA = (57) : Temporal variations in one or more characteristics measured from 800 a cell-free DNA sample are used to estimate a gestational age of a fetus. Exam- — 810 ple characteristics include the methylation level measured from the cell-free DNA sample, size of DNA fragments measured from the cell-free DNA sample (e.g., = Arialysits ere-free DNA isoleasks Pisa a Siuttple to determine whether the cell-free DNA molecule is proportion of fetal-derived DNA fragments longer than a specified size), and end- = = — Methylated At one mmotn Sites. ing patterns of the DNA fragments align to a reference genome. =8ID Determine arespectivertumber of cell-free DNA —_ oloteculea thar aremedaylated at each of the one ormore sites = = = — Calculate ameasund methylation level of cell-free DNA = Moleattlea in the sample = Obtain ate or more tatibtathin data pokis Shag speedy a gestational age corresponding to a calibration methylation level 11 850 in Compare the measured metbylation Level to a calibratima medtgladon level of at kart one calibration data points IN 11 © g6 1 N Estimate a gestational age of the fetus based owthe Ir----- comparison © 1-1 Fig. 8 0 N O [Continued on next page] WO 2018/072705 Al MIDEDIMOMMIDIREIHNIFIHMEIIMERVOIMIE TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG). Published: — with international search report (Art. 21(3))
GESTATIONAL AGE ASSESSMENT BY METHYLATION AND SIZE PROFILING OF MATERNAL PLASMA DNA