4. Characteristics of included studies by type of gNIPT.

Study ID	Target condition(s)	Study design and participants	Prior risk	Index test details	Cutpoint	Reference standard	Comparator
MPSS
Alberti 2015	T21	Case‐control study (1:2) from a prospective cohort 976 singleton pregnancies enrolled, 183 were analysed	High risk	Illumina HiSeq 2000 sequencer without multiplexing In‐house test FF measured	Z score of 3	Fetal karyotypea
Benachi 2015	T21, T18, T13	Blinded retrospective study 900 singleton or twin pregnancies enrolled, 886 were analysed	High risk	Illumina v3 flow‐cell on a HiSeq 1500 sequencer in 12‐plex Commercial ‐ Laboratoire CERBA FF measured	Z score of 3 for T21; 3.95 for T18 and T13	Fetal karyotype or neonatal clinical examination
Bianchi 2012	T21, T18, T13, 45,X, 47,XXX, 47,XXY, 47,XYY	Nested case‐control study (1:4) from a prospective cohort (MELISSA) 2882 singleton pregnancies enrolled, 503 for T21, 502 for T18, 501 for T13 and 489 for 45,X were analysed	High risk	Illumina HiSeq 2000 sequencer in 6‐plex Commercial test ‐ Verinata FF measured	Different cutpoints used for autosomes and SCAb	Fetal karyotype
Bianchi 2013	T21, T18, T13, 45,X	Retrospective study from stored plasma 2882 singleton pregnancies enrolled, 113 were analysed	High risk	Illumina TrueSeq 3.0 sequencing chemistry Commercial test ‐ Verinata	Different cutpoints used for autosomes and SCAb	Fetal karyotype
Bianchi 2014a	T21, T18, T13	Blinded prospective cohort study 2052 singleton pregnancies enrolled, 1952 for T21 and T18, and 1914 for T13 were analysed	High, low and without prior risk	Illumina HiSeq 2000 in 8‐plex Commercial ‐ verifi® prenatal test FF measured	NCV of 4; resequenced if NCV is between 3 and 4	Fetal or postnatal karyotype, neonatal clinical examination or medical record from birth	Standard screening (T21 only with mixed cutpoints) which include first‐trimester combined test or a second‐trimester result (quadruple, serum integrated, fully integrated, or sequential).
Bijok 2014	T21, T18, T13	Prospective cohort study 10 singleton pregnancies enrolled, 9 were analysed	High risk	IIIumina Genome Analyzer IIx or HiSeq 2000 sequencer in multiplex Commercial ‐ NIFTY™ test, BGI‐Shenzhen FF measured	NR	Fetal karyotype
Canick 2012	T21, T18, T13	Case‐control study 4664 pregnant women enrolled, 27 multifetal pregnancies were analysed	High risk	Illumina HiSeq 2000 sequencer in 4‐plex Commercial test ‐ Sequenom, Inc. FF measured	Z score of 3	Fetal karyotype
Chen 2011	T18, T13	Nested case‐control study from prospective and retrospective cohorts 392 singleton pregnancies enrolled, 289 were analysed	High risk	Illumina Genome Analyzer IIx in 2‐plex Commercial test ‐ Sequenom, Inc.	Z score of 3	Fetal karyotype
Chiu 2011	T21	Blinded case‐control study (1:5) from prospective and retrospective cohorts 824 singleton pregnancies enrolled, 753 were analysed by 8‐plex method and 314 by 2‐plex method	Mostly high (> 1/300) and some intermediate risk (between 1/300 and 1/1000)	Illumina Genome Analyzer II in 8‐plex and 2‐plex Commercial test ‐ Sequenom, Inc. FF measured	Z score of 3	Fetal karyotype
Ehrich 2011	T21	Blinded case‐control study (1:11) from prospective cohort 480 pregnant women enrolled, 449 were analysed	High risk	Illumina Genome Analyzer IIx sequencer in 4‐plex Commercial test ‐ Sequenom, Inc. FF measured	Z score of 2.5	Fetal karyotype
Fiorentino 2016	T21, T18, T13	Blinded prospective cohort study 7103 singleton pregnancies enrolled, 7082 were analysed	Mostly high risk and without prior risk	Illumina HiSeq 2500 sequencer in 15‐plex, SAFeR™ algorithm. Commercial ‐ Genoma's prenatal test FF measured	NCV of 4; aneuploidy suspected if NCV is between 3 and 4	Fetal karyotype or neonatal clinical examination
Hou 2012	T21, T18, T13, 45,X, 47,XXX, 47,XXY, 47,XYY	Prospective cohort study 308 singleton pregnancies enrolled, 205 were analysed	High risk	IIIumina HiSeq 2000 sequencer Commercial test ‐ BGI‐Shenzhen	NR	Fetal karyotype
Huang 2014	T21, T18	Blinded prospective cohort study 189 twin pregnancies enrolled, 189 were analysed	High risk	IIIumina Genome Analyzer IIx or HiSeq 2000 sequencer Commercial test ‐ BGI‐Shenzhen	L score of 1 and t score of 2.5 including warning zone	Fetal karyotype
Jeon 2014	T21, T18	Prospective cohort study 155 singleton pregnancies enrolled, 155 were analysed	High risk	Ion Torrent PGM or HiSeq 2000 sequencers, 10 samples per Chip Commercial test ‐ Genome Care	Z score of 2.566 for T21; 2.459 for T18.	Fetal karyotype
Jiang 2012	T21, T18, T13, 45,X, 47,XXY, 47, XYY	Prospective cohort study 903 pregnant women enrolled, 903 were analysed	High risk	IIIumina Genome Analyzer IIx or HiSeq 2000 sequencer in multiplex Commercial ‐ NIFTY™ test, BGI‐Shenzhen FF measured	Different cutpoints used for autosomes and SCAb	Fetal karyotype
Johansen 2016	T21, T18, T13	Prospective cohort study 375 singleton pregnancies enrolled, 173 were analysed	High risk	Ion Proton™ sequencer in 5‐plex In‐house test FF measured	Z score of 4 (unclassified if Z score is between 3 and 4) and WISECONDOR of 1%	Fetal karyotype
Ke 2015	T21, T18, T13	Prospective cohort study 2340 singleton pregnancies enrolled, 2340 were analysed	High risk	High throughput sequencing platform Commercial test ‐ BGI‐Shenzhen	T score of 3	Fetal karyotype or newborn outcome
Kim 2016	T21	Blinded prospective cohort study 101 pregnant women enrolled, 101 were analysed	High risk	Ion Proton™ sequencer in multiplex Commercial test ‐ Genome Care	Z score of 2.10 for Ion Proton™	Fetal karyotype
Lau 2012	T21, T18, T13, 45,X, 47,XXX, 47,XXY, 47,XYY	Blinded prospective cohort study 108 singleton pregnancies enrolled, 108 were analysed	Mostly high risk	IIIumina HiSeq 2000 sequencers in 12‐plex Commercial ‐ NIFTY™ test, BGI‐Shenzhen	Different cutpoints used for autosomes and SCAb	Fetal karyotype
Lee 2015	T21, T18, T13 and SCA (no case found)	Blinded prospective cohort study 93 singleton and multifetal pregnancies enrolled, 92 were analysed	High risk	Illumina MiSeq sequencer in 12‐plex or NextSeq sequencer in 96‐plex Commercial test ‐ MomGuard™, LabGenomics FF measured	Z score of 4 (intermediate risk if Z score is between 2.5 and 4) for T21 and T18; 2.8 for T13 (intermediate risk if Z score is between 1.9 and 2.8)	Fetal or neonatal karyotype
Lefkowitz 2016	T21, T18, T13, 45,X, 47,XXX, 47,XXY, 47,XYY	Retrospective cohort, blinded case‐control study 5321 pregnant women enrolled but 1222 were selected and 1166 were analysed	High risk	IIIumina HiSeq 2000 sequencer in 6‐plex or uniplex Commercial test ‐ Sequenom, Inc. FF measured	Different cutpoints used for autosomes and SCAb	Fetal karyotype
Liang 2013	T21, T18, T13, 45,X, 47,XXX, 47,XXY, 47,XYY	Blinded prospective cohort study 435 singleton and twin pregnancies enrolled, 412 were analysed	High risk	Illumina HiSeq 2000 sequencer in 8‐plex or 12‐plex Commercial test ‐ Berry Genomics Co. Ltd. FF measured	Different cutpoints used for autosomes and SCAb	Fetal karyotype
Liu 2012	T21, T18, T13, 45,X, 47,XXX, 47,XXY, 47,XYY	Prospective cohort study 153 pregnant women enrolled, 153 were analysed	High risk	Illumina HiSeq sequencer in multiplex.	Z score of 3	Fetal karyotype
Ma 2016	T21, T18, T13	Blinded retrospective (archived samples) and prospective cohorts study 10,598 singleton pregnancies enrolled, 10,579 were analysed	High and low risk	Sequencing on BGISEQ‐1000 in 16 or 24‐plex Commercial test ‐ BGI‐Shenzhen	Z score of 3	Fetal karyotype or postnatal follow‐up
Mazloom 2013	45,X, 47,XXX, 47,XXY, 47,XYY	Blinded prospective cohort study 1975 singleton pregnancies enrolled, 411 samples from the validation set were analysed	High risk	Illumina v3 flow‐cell on a HiSeq 2000 sequencer in 12‐plex Laboratory test development by Sequenom, Inc. FF measured	Different cutpoints used for the four SCAb	Fetal karyotype
Palomaki 2012	T21, T18, T13	Nested case‐control study (1:3) 4664 pregnant women enrolled but 1988 singleton pregnancies were selected and 1971 were analysed	High risk	Illumina HiSeq 2000 sequencer in 4‐plex Commercial test ‐ Sequenom, Inc. FF measured	Z score of 3 for T21; 3.88 for T18; 7.17 for T13	Fetal karyotype
Papageorghiou 2016a	T21, T18, T13	Retrospective cohort, case‐control study (1:9) 442 singleton and twin pregnancies enrolled, 426 singleton pregnancies were analysed	High risk	Ion Proton™ sequencer in 8‐plex Commercial ‐ IONA® test, Premaitha Health (public limited company in UK) FF measured	Likelihood ratio of 1 and maternal age‐adjusted probability risk score	Fetal karyotype or medical record from birth
Papageorghiou 2016b	T21, T18, T13	Retrospective cohort, case‐control study (1:9) 442 singleton and twin pregnancies enrolled, 11 twin pregnancies were analysed	High risk	Ion Proton™ sequencer in 8‐plex Commercial ‐ IONA® test, Premaitha Health (public limited company in UK) FF measured	Likelihood ratio of 1 and maternal age‐adjusted probability risk score	Fetal karyotype or medical record from birth
Poon 2016	T21, T18, T13	Retrospective cohort, blinded nested case‐control study 242 singleton pregnancies enrolled, 241 were analysed	High risk	Ion Proton™ sequencer, IONA® software algorithm Commercial ‐ IONA® test, Premaitha Health (public limited company in UK) FF measured	NR (authors used the same gNIPT than Papageorghiou 2016a)	Fetal karyotype
Porreco 2014	T21, T18, T13, 45,X, 47,XXX, 47,XXY, 47,XYY	Blinded prospective cohort study 4170 singleton pregnancies enrolled, 3322 for autosomes, 3278 for 45,X and 47,XXX and 3201 for 47,XXY and 47,XYY were analysed	High risk	Illumina HiSeq 2000 sequencer in 12‐plex Commercial test ‐ Sequenom, Inc. FF measured	Different cutpoints used for autosomes and SCAb	Fetal karyotype or medical record from birth
Sehnert 2011	T21, T18, T13, 45,X	Retrospective (archived samples) cohort study 1014 singleton and multifetal pregnancies enrolled but only 47 singleton pregnancies in the test set were analysed in this review.	High risk	IIIumina Genome Analyzer IIx sequencer in uniplex Commercial test ‐ Verinata	Different cutpoints used for autosomes and SCAb	Fetal karyotype
Shaw 2014	T21, T18, T13, 45,X, 47, XXX, 47,XXY, 47,XYY	Prospective cohort study 201 singleton and multifetal pregnancies enrolled, 200 were analysed	High and low risk	Illumina v2 HiSeq 2000 sequencer in 12‐plex Commercial test ‐ Berry Genomics Co. Ltd.	Different cutpoints used for autosomes and SCAb	Fetal karyotype or medical record from birth
Song 2013	T21, T18, T13, 45,X, 47,XXX, 47, XXY, 47,XYY (SCA data not shown in this review)	Blinded prospective cohort study 1916 singleton pregnancies enrolled, 1741 were analysed	Without prior risk	Illumina v2 HiSeq2000 in 12‐plex Commercial test‐ Berry Genomics Co. Ltd.	Z score of 3	Fetal or postnatal karyotype or medical record from birth	Triple test for T21 and T18 (cutpoint of 1 in 270).
Song 2015	T21, T18, T13, 45,X, 47,XXX, 47,XYY	Blinded prospective cohort study 213 singleton pregnancies enrolled, 204 were analysed	High risk	Illumina v2 HiSeq 2000 sequencer in 12‐plex Commercial test ‐ Berry Genomics Co. Ltd. FF measured	Z score of 3	Fetal karyotype or neonatal clinical examination or both
Stumm 2014	T21, T18, T13	Prospective cohort, blinded study for T21 and unblinded for T18 and T13 522 singleton pregnancies enrolled, 472 were analysed	High risk	Illumina HiSeq 2000 sequencer in 12‐plex (DAP.21 algorithm without CG correction) Commercial test ‐ LifeCodexx AG FF measured	MAD‐based Z score of 3 for T21; 3.2 for T18; 3.9 for T13	Fetal karyotype
Sukhikh 2015	T21, T18, T13, 45,X	Prospective cohort study 200 pregnant women enrolled, 200 were analysed	High risk	Ion Proton™ sequencer In‐house test	T score of 5 for T21 and T18; 4 for T13; 0.04 Chrom. X and 0.04 Chrom. Y for 45,X	Fetal karyotype
Sung‐Hee 2015	T21, T18, T13, 45,X, 47,XXX, 47,XXY, 47,XYY	Retrospective study 918 singleton pregnancies enrolled, 901 were analysed	High risk	IIIumina Genome Analyzer IIx or HiSeq 2000 sequencer in 12‐plex Commercial ‐ NIFTY™ test, BGI‐Shenzhen FF measured	L score of 1 and t score of 2.5	Fetal karyotype or medical record from birth
Tynan 2016	T21, T18, T13	Blinded retrospective cohort study 1100 singleton pregnancies enrolled, 1048 were analysed	High and without prior risk	Illumina HiSeq 2000 or HiSeq 2500 sequencers in multiplex Commercial ‐ VisibiliT™ test, Sequenom, Inc. FF measured	risk score of 1%	Fetal karyotype or medical record from birth
Wang 2014	T21, T18, T13, 45,X	Prospective cohort study 136 singleton pregnancies enrolled, 136 were analysed	High risk	Illumina HiSeq 2000 sequencer Commercial ‐ NIFTY™ test, BGI‐Shenzhen	NR	Fetal or neonatal karyotype or clinical examination at 42 days after birth or both
Wang 2015a	T21, T18, T13, 45,X, 47,XXX, 47,XXY, 47,XYY	Prospective cohort study 917 pregnant women enrolled, 917 were analysed	High risk	Illumina v2 HiSeq 2000 flow cell on a HiSeq sequencer Commercial test ‐ Berry Genomics Co. Ltd	Z score of 3 for T21, T18 and T13; ‐3 for Chrom. X and 3 for Chrom. Y for sex Chrom. classification.	Fetal karyotype or clinical follow‐up to 6 months from birth
Yao 2014	T21, T18, T13 and SCA (SCA data not shown in this review)	Retrospective study 5950 singleton pregnancies enrolled, 5530 were analysed	High, low and without prior risk	IIIumina Genome Analyzer IIx or HiSeq 2000 sequencer in 12‐plex Commercial ‐ NIFTY™ test, BGI‐Shenzhen FF measured	Different cutpoints used for autosomes and SCAb	Fetal karyotype or clinical follow‐up
Zhang 2016	T21, T18, 45,X, 47,XXX (SCA data not shown in this review)	Blinded prospective cohort study 87 singleton pregnancies enrolled, 87 were analysed	High risk	Illumina HiSeq 2000 sequencer in 12‐plex Commercial test ‐ Berry Genomics Co. Ltd.	Z score of 3 for T21 (no other cutpoint reported)	Fetal or neonatal karyotype or neonatal clinical examination
Zhou 2014a	T21, T18, T13	Blinded prospective cohort study 306 singleton pregnancies enrolled, 301 were analysed	High, low and without prior risk	IIIumina Genome Analyzer IIx or HiSeq 2000 sequencer in 12‐plex Commercial ‐ NIFTY™ test, BGI‐Shenzhen FF measured	L score of 1 and t score of 2.5	Fetal or neonatal karyotype or birth outcome
Zhou 2014b	T21, T18, T13	Blinded prospective cohort study 7705 singleton pregnancies enrolled, 3950 were analysed	High, low and without prior risk	IIIumina Genome Analyzer IIx or HiSeq 2000 sequencer in 12‐plex Commercial ‐ NIFTY™ test, BGI‐Shenzhen FF measured	L score of 1 and t score of 2.5	Fetal or neonatal karyotype or birth outcome
TMPS
Ashoor 2012	T21, T18	Nested case‐control study (1:3) from a prospective cohort 400 singleton pregnancies enrolled, 397 were analysed	High risk	DANSR assay (FORTE algorithm), Illumina HiSeq 2000 in 96‐plex Commercial ‐ Harmony™ prenatal test, Ariosa Diagnostics, Inc.	NR (usually Harmony™ prenatal test uses FORTE risk score of 1%)	Fetal karyotype
Ashoor 2013	T13	Blinded prospective cohort study 2167 singleton pregnancies enrolled, 1949 were analysed	High and low risk	DANSR assay (FORTE algorithm), Illumina HiSeq 2000 in 96‐plex Commercial ‐ Harmony™ prenatal test, Ariosa Diagnostics, Inc. FF measured	FORTE risk score of 1%	Fetal karyotype or neonatal clinical examination
Bevilacqua 2015	T21, T18, T13	Prospective cohort study 515 multifetal pregnancies enrolled, 340 were analysed Women with singleton pregnancies were excluded (incomplete 2 x 2 table).	High and without prior risk	DANSR assay (FORTE algorithm), Illumina HiSeq 2000 in 96‐plex Commercial ‐ Harmony™ prenatal test, Ariosa Diagnostics, Inc. FF measured	NR (usually Harmony™ prenatal test uses FORTE risk score of 1%)	Fetal or neonatal karyotype
Comas 2015	T21, T18, T13, 45,X, 47,XXX, 47, XXY, 47,XYY (SCA data not shown in this review)	Blinded prospective cohort study 333 singleton pregnancies enrolled, 312 were analysed	High and without prior risk	DANSR assay (FORTE algorithm) or SNP‐based method Commercial ‐ Panorama™ test, Natera, Inc. or Harmony™ prenatal test, Ariosa Diagnostics, Inc. FF measured	Harmony™ prenatal test: NR (usually Harmony™ prenatal test uses FORTE risk score of 1%) Panorama™ test: NR	Fetal karyotype or neonatal clinical examination
del Mar Gil 2014	T21, T18, T13	Retrospective cohort study 207 multifetal pregnancies enrolled, 192 twin pregnancies were analysed	Without prior risk	DANSR assay (FORTE algorithm), Illumina HiSeq 2000 in 96‐plex Commercial ‐ Harmony™ prenatal test, Ariosa Diagnostics, Inc. FF measured	NR (usually Harmony™ prenatal test uses FORTE risk score of 1%)	Fetal karyotype
Gil 2016	T21, T18, T13	Prospective cohort study 11,692 singleton pregnancies enrolled, 3633 were analysed	High and intermediate riskc	DANSR assay (usually with FORTE algorithm) Commercial ‐ Harmony™ prenatal test, Ariosa Diagnostics, Inc.	NR (usually Harmony™ prenatal test uses FORTE risk score of 1%)	Fetal or postnatal karyotype or neonatal clinical examination
Hall 2014	T13	Case‐control study (1:3)/1000 singleton pregnancies enrolled, 64 were analysed.	High risk	SNP‐based method (NATUS algorithm), IIIumina Genome Analyzer IIx or HiSeq sequencer, 11,000 or 19,488‐plex targeted PCR Commercial ‐ Natera's prenatal test FF measured	NR	Fetal karyotype or genetic testing of cord blood, buccal, saliva or products of conception
Hooks 2014	45,X, 47,XXX, 47, XXY, 47,XYY	Case‐control study from archived samples 432 singleton pregnancies enrolled, 414 were analysed	High risk	DANSR assay (FORTE algorithm), Illumina HiSeq 2000 in 96‐plex Commercial ‐ Harmony™ prenatal test, Ariosa Diagnostics, Inc. FF measured	NR (usually Harmony™ prenatal test uses FORTE risk score of 1%)	Fetal karyotype
Jackson 2014	T21, T18, T13	Prospective cohort study 1228 pregnant women enrolled, 1161 were analysed	High and low risk	DANSR assay (FORTE algorithm) Commercial ‐ Harmony™ prenatal test, Ariosa Diagnostics, Inc.	NR (usually Harmony™ prenatal test uses FORTE risk score of 1%)	Fetal karyotype or medical record from birth
Korostelev 2014	T21, T18, T13, 45,X, 47,XXX, 47, XXY, 47,XYY	Prospective cohort study 1968 singleton pregnancies enrolled, 685 were analysed	High and without prior risk	SNP‐based method (NATUS algorithm), IIIumina Genome Analyzer IIx or HiSeq sequencer, > 19,000‐plex targeted PCR Commercial ‐ Natera's prenatal test FF measured	NR	Fetal karyotype or medical record from birth
Nicolaides 2012	T21, T18	Retrospective study from archived plasma 2230 singleton pregnancies enrolled, 1949 were analysed	Without prior risk	DANSR assay (usually with FORTE algorithm) Commercial ‐ Harmony™ prenatal test, Ariosa Diagnostics, Inc. FF measured	Risk score of 1%	Fetal karyotype or neonatal clinical examination	First‐trimester combined test (cutpoint of 1 in 150).
Nicolaides 2013	T21, T18, T13, 45,X, 47,XXX, 47,XXY, 47,XYY	Blinded prospective cohort study 242 singleton pregnancies enrolled, 229 were analysed	High risk	SNP‐based method (NATUS algorithm), IIIumina Genome Analyzer IIx or HiSeq sequencer, 19,488‐plex targeted PCR Commercial ‐ Natera's prenatal test FF measured	NR	Fetal karyotype
Nicolaides 2014a	45,X, 47,XXX, 47,XXY, 47,XYY	Case‐control study (archived samples) 177 singleton pregnancies enrolled, 172 were analysed	High risk	DANSR assay (FORTE algorithm), Illumina HiSeq 2000 in 96‐plex Commercial ‐ Harmony™ prenatal test FF measured	FORTE risk score of 1%	Fetal karyotype
Norton 2012	T21, T18	Blinded prospective cohort study 4002 singleton pregnancies enrolled, 3080 were analysed	High risk	DANSR assay (FORTE algorithm), Illumina HiSeq 2000 in 96‐plex Commercial test‐ Ariosa Diagnostics, Inc. FF measured	FORTE risk score of 1%	Fetal karyotype
Norton 2015	T21, T18, T13	Blinded prospective cohort study 18,955 singleton pregnancies enrolled, 15,841 were analysed	Without prior risk	DANSR assay (FORTE algorithm) Commercial ‐ Harmony™ prenatal test, Ariosa Diagnostics, Inc. FF measured	NR (usually Harmony™ prenatal test uses FORTE risk score of 1%)	Fetal or postnatal karyotype, neonatal clinical examination or medical record from birth	First‐trimester combined test (cutpoint of 1 in 270 for T21 and 1 in 150 for T18 and T13).
Pergament 2014	T21, T18, T13, 45,X	Blinded prospective cohort study 1064 singleton pregnancies enrolled, 963 were analysed	High and low risk	SNP‐based method (NATUS algorithm), IIIumina Genome Analyzer IIx or HiSeq sequencer, 19,488‐plex targeted PCR Commercial ‐ Natera's prenatal test FF measured	NR	Fetal karyotype or genetic testing of cord blood, buccal, saliva or products of conception or birth outcome
Persico 2016	T21, T18, 45,X, 47,XXX, 47,XXY, 47,XYY	Blinded prospective cohort study 259 singleton pregnancies enrolled, 249 were analysed	High risk	SNP‐based method (NATUS algorithm), IIIumina Genome Analyzer IIx or HiSeq sequencer, 19,488‐plex targeted PCR Commercial ‐ Natera's prenatal test FF measured	Risk score of 1%	Fetal karyotype
Quezada 2015	T21, T18, T13	Prospective cohort study 2905 singleton pregnancies enrolled, 2785 were analysed	Without prior risk	DANSR assay (FORTE algorithm) Commercial ‐ Harmony™ prenatal test FF measured	NR (usually Harmony™ prenatal test uses FORTE risk score of 1%)	Fetal or postnatal karyotype, neonatal clinical examination or medical record from birth	First‐trimester combined test (cutpoint of 1 in 100 for T21).
Samango‐Sprouse 2013	45,X, 47,XXX, 47,XXY, 47,XYY	Blinded prospective cohort study 201 singleton pregnancies (with known SCA and euploid pregnancies) enrolled, 186 were analysed	High and low risk	SNP‐based method (NATUS algorithm), IIIumina HiSeq sequencer, 19,488‐plex targeted PCR Commercial ‐ Natera's prenatal test FF measured	NR	Fetal karyotype or genetic testing of cord blood, buccal, saliva or products of conception
Sparks 2012a	T21, T18	Case‐control study from a prospective cohort 338 singleton pregnancies enrolled, 167 were analysed	High risk	DANSR assay (FORTE algorithm), Illumina HiSeq 2000 in 96‐plex Commercial test‐ Ariosa Diagnostics, Inc. FF measured	NR	Fetal karyotype
Verweij 2013	T21	Blinded prospective cohort study 595 singleton pregnancies enrolled, 504 were analysed	High risk	DANSR assay (FORTE algorithm), Illumina HiSeq 2000 in 96‐plex Commercial test‐ Ariosa Diagnostics, Inc. FF measured	FORTE risk score of 1%	Fetal karyotype

45,X: Turner syndrome, 47,XXX: triple X syndrome, 47,XXY: Klinefelter syndrome, DANSR: digital analysis of selected regions, FF: fetal fraction DNA, FORTE: fetal‐fraction optimised risk of trisomy evaluation, MAD: Median absolute deviation, MPSS: massively parallel shotgun sequencing, NATUS: Next‐generation Aneuploidy Test Using SNPs, NCV: normalised chromosome value, SCA: sex chromosome aneuploidy, SNP: single‐nucleotide polymorphism,TMPS: targeted massively parallel sequencing, T21: trisomy 21, T18: trisomy 18 and T13: trisomy 13.

^aFetal karyotype include traditional banding techniques, spectral karyotype, fluorescence in situ hybridisation, array comparative genomic hybridisation or quantitative fluorescence polymerase chain reaction.

^bDifferent cutpoints used for autosomes or SCA as follows:

Bianchi 2012: NCV of 4 (aneuploidy suspected if NCV is between 2.5 and 4) for T21, T18, and T13; NCV for Chrom. X of ‐4 and NCV for Chrom. Y of 2.5 for 45,X; NCV for Chrom. X of 4 and NCV for Chrom. Y of 2.5 for 47,XXX; NCV for Chrom. X between ‐2.5 and 2.5 and NCV for Chrom. Y > 33 for 47,XXY; NCV for Chrom. X of ‐4 and NCV for Chrom. Y of 4 for 47,XYY with NCV for Chrom. Y is two times greater than expected NCV Chrom. X.

Bianchi 2013: NCV of 4 (aneuploidy suspected if NCV is between 3 and 4) for T21, T18, and T13; NCV for Chrom. X of ‐3 and NCV for Chrom. Y of 3 for 45,X.

Jiang 2012: t score of 3 and logarithmic LR of 1 for T21, T18 and T13; if female fetus, t score of ‐2.5 for 45,X and 47,XXX; t score of 2.5 combined with estimation of fetal ccfDNA concentration by Chrom. X and Y independently for 47,XXY and 47,XYY.

Lau 2012: Z score of 3 for T21, T18 and T13; if female fetus, Z score for Chrom. X of ‐3 for 45,X; if female fetus, Z score for Chrom. X of 3 for 47,XXX; if male fetus, Z score for Chrom. Y of 3 for 47,XXY.

Lefkowitz 2016: Z score of 3 for T21; Z score of 3.95 for T18 and T13; Z scores for SCA see Mazloom 2013.

Liang 2013: Z score of 3 for T21; 5.91 for T18; 5.72 for T13; ± 2.91 for Chrom. X and ± 3 for Chrom. Y for sex chromosome classification.

Mazloom 2013: Z score of 3.5 for 47,XXX (non‐reportable regions between 2.5 and 3.5); Z score of ‐3.5 for 45,X (non‐reportable regions between ‐2.5 and ‐3.5); Z score of ‐3.5 for 47,XYY with Chrom. Y representation; between ‐3.5 and 3.5 for 47,XXY with Chrom. Y representation.

Porreco 2014: Z score of 3 for T21; Z score of 3,95 for T18 and T13; Z score of 3.5 for 47,XXX (non‐reportable regions between 2.5 and 3.5); Z score of ‐3.5 for 45,X (non‐reportable regions between ‐2.5 and ‐3.5); Z score of ‐3.5 for 47,XYY with Chrom. Y representation; Z score between ‐3.5 and 3.5 for 47,XXY with Chrom. Y representation.

Sehnert 2011: NCV of 4 (unclassified if NCV is between 2.5 and 4) for T21, T18, and T13; NCV for Chrom. Y of ‐2.0 SDs from the mean of male samples and NCV for Chrom. X of ‐3.0 SDs from the mean of female samples for sex chromosome classification.

Shaw 2014: Z score of 3 for T21, T18, and T13; Z score of ‐3 for Chrom. X and 3 for Chrom. Y for sex chromosome classification.

Yao 2014: T score of 2.5 for T21, T18 and T13; if female fetus, T score for Chrom. X of ‐2.5 for 45,X and 2.5 for 47,XXX; if male fetus, T score for Chrom. X of 2.5 combined with estimation of fetal ccfDNA concentration by Chrom. X (expected value of zero) for 47,XXY; if male fetus, T score for Chrom. X of 2.5 and R‐value (the ratio of the fetal DNA fraction estimated by chromosome Y to that estimated by chromosome X) between 1.8 and 2.2 for 47,XYY.

^cPregnant women with a first‐trimester combined test selected for their risk of fetal aneuploidy (cutpoint of 1 in 100 for high risk and 1 in 101 to 1 in 2500 for intermediate risk).