What is. PrenatalSAFE Karyo test is the most technologically advanced genome-wide NIPT

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What is PrenatalSAFE Karyo test is the most technologically advanced genome-wide NIPT Through cfdna analysis from maternal plasma, PrenatalSAFE Karyo detects: Aneuploidies structural chromosomal aberrations (deletions or duplications) across the fetal genome providing karyotype-level insight.

the next level in noninvasive prenatal testing PrenatalSafe Karyo analyzes every chromosome in the genome. Unlike any other noninvasive prenatal test available to date, it offers a level of information previously only available from a fetal karyotype analysis, performed with invasive prenatal diagnosis procedures (amniocentesis and CVS)

NIPT uses cell-free DNA (cfdna) Maternal blood sample Maternal cfdna Fetal cfdna

Whole-genome sequencing technology cfdna extraction Fetal Fraction determination DNA Sequences counting

2 levels of genome-wide screening It can also test for 9 common microdeletion syndromes Microdeletion Syndrome Prevalence DiGeorge syndrome (22q11.2) 1 in 4,000 1p36 deletion syndrome 1 in 4,000 to 1 in 10,000 Angelman syndrome (15q11.2) 1 in 12,000 Prader-Willi syndrome (15q11.2) 1 in 10,000 to 1 in 25,000 Cri du Chat syndrome (5p15.3) 1 in 20,000 to 1 in 50,000 Wolf-Hirschhorn syndrome (4p16.3) 1 in 50,000 Langer-Giedion syndrome (8q24) 1/200.000 Jacobsen syndrome (11q23) 1/100.000 Smith-Magenis syndrome (17p11.2) 1/15.000-1/25.000

Detection rate

Detection rate PrenatalSAFE Karyo Plus test identifies 95.5% of chromosomal anomalies prenatally detected and 99.1% of those anomalies observed at birth, achieving a level of detection rate very closed to the traditional fetal karyotyping (96.9%), obtained with invasive, prenatal diagnostic techniques.

Conventional Prenatal Screening vs Detection Rates for Trisomy 21 Quadruple Screen 1st Trimester Blood Screen + NT Serum Integrated False Positive Rate (FPR): 5% Integrated Screen ACOG Practice Bulletin No. 77, January 2007

Fetal Karyotyping vs Analyzes every chromosome Traditional Karyotyping Detects chromosomal gains or losses >7 Mb Detects chromosome gains or losses >10 Mb Requires an invasive procedure Detects unbalanced translocations Detects segmental deletions/duplications Detects mosaic trisomies Detects marker chromosomes Detects microdeletion syndromes Detects triploidy Considered diagnostic

Pre-clinical Validation performance Trisomy 21 (n=1419) Trisomy 18 (n=1419) Trisomy 13 (n=1419) SCA (n=1419) CNV (n=1419) True Positive 100 31 14 36 37 False Positive 0 0 0 0 0 True Negative 1319 1388 1405 1383 1382 False Negative 0 0 0 0 0 Sensitivity (95% CI) Specificity (95% CI) PPV (95% CI) NPV (95% CI) (96.38% - 100.00%) (99.72% - 100.00%) (96.38% - 100.00%) (99.72% - 100.00%) (88.78% - 100.00%) (99.73% - 100.00%) (88.78% - 100.00%) (99.73% - 100.00%) (76.84% - 100.00%) (99.74% - 100.00%) (76.84% - 100.00%) (99.74% - 100.00%) (90.26% to 100.00%) (99.73% to 100.00%) (90.26% to 100.00%) (99.73% to 100.00%) (90.51% to 100.00%) (99.73% to 100.00%) (90.51% to 100.00%) (99.73% to 100.00%) PPV: Positive Predictive Value; NPV: Negative Predictive Value; SCA: Sex Chromosomes Aneuploidy. CNV: Copy Number Variation Fiorentino et al., EJHG conference 2016; ISPD conference 2016

Performance Standard protocol Follow-up March 2016 PPV: Positive Predictive Value; NPV: Negative Predictive Value; SCA: Sex Chromosomes Aneuploidy

Performance FAST protocol Follow-up March 2016 PPV: Positive Predictive Value; NPV: Negative Predictive Value; SCA: Sex Chromosomes Aneuploidy

Published validation studies

This test is intended for patients at 10 weeks or greater gestation who meet any of the following criteria: Maternal age-related risks (e35 years) Positive results on maternal serum screening Abnormal ultrasound finding(s) Prior pregnancy with aneuploidy Parental translocation Low risk pregnancies Patients wanting early, accurate testing and are at average risk of aneuploidy Which Patients Should Be Offered?

pregnancy characteristics The test is suitable for both single and twin pregnancies. It can be performed in patients whose pregnancies have been achieved by IVF techniques, including pregnancies with egg donation.

Free services supplied

Case study

Case Study 1 31 y.o. patient 12^ weeks gestation 5 Negative Result 20^ week gestation Fetal abnormal ultrasound findings (cerebellar ipoplasia; Ventriculomegaly ) Amniocentesis Reassessment of the NIPT data with

Chr. 5 Detection of a deletion 5p confirmed by array-cgh Chr. 5 del5p15.33p13.2 34 Mb Amniotic Fluid cfdna Array-CGH PrenatalSafe Karyo

Case Study 2 32 y.o. patient 13^ weeks gestation 5 Negative Result 21^ weeks gestation Multiple fetal abnormal ultrasound findings Reassessment of the NIPT data with Amniocentesis

Chr. 18 Identification of 2 deletions (18p / 18q) confirmed by array-cgh Chr. 18 Del18p11.32-p11.31 3.8 Mb Amniotic Fluid cfdna Del18q21.32-q23 21.3 Mb Array-CGH PrenatalSafe Karyo

Case Study 3 32 y.o patient Carrier of a reciprocal translocation 46,XX,t(7;9)(p15;q22) 11^ weeks gestation Chr. 7 Chr. 9 Abnormal Result detected fetal karyotype with an unbalanced translocation Villocentesis Results confirmed

Detected a fetal karyotype with an unbalanced translocation confirmed by array-cgh Dup7p22.3p21.2 15 Mb Dup9p24.3-q31.1 107.5 Mb Array-CGH PrenatalSafe Karyo Array-CGH PrenatalSafe Karyo CVS cfdna CVS cfdna

Case Study 4 35 y.o patient 13^ weeks gestation Amniocentesis (traditional karyotyping) Negative Result (Fetale karyotype 46, XY) (Suspected DiGeorge syndrome) 20^ weeks gestation Fetal abnormal ultrasound findings Identified deletion 22q11.2 (compatible with DiGeorge syndrome) Amniocentesis

Plus Identification of 22q11.2 deletion confirmed by Array-CGH Chr.22 del22q11.21 DiGeorge Syndrome 4 Mb Amniotic Fluid cfdna Array-CGH PrenatalSafe Karyo

Case Study 5 Chr. 13 Chr. 20 33 y.o. patient (She was not aware to be carrier of a chromosomal translocation) 12^ weeks gestation Abnormal Result detected fetal karyotype with an unbalanced translocation Villocentesis Paziente portatrice di traslocazione reciproca 46,XX,t(13;20)(q33;q13.3) Results confirmed

Detected a fetal karyotype with an unbalanced translocation confirmed by array-cgh Del13q33.1q34 11.4 Mb Dup20q13.33 1.9 Mb Chr. 13 Chr. 20 PrenatalSafe Karyo (cfdna) Array-CGH (CVS) PrenatalSafe Karyo (cfdna) Array-CGH (CVS)

Case Study 6 40 y.o. patient 13^ weeks gestation NIPT (5 chromosomes screening) Positive Result High risk for Trisomy 21 Test performed by a different lab Trisomy 21 not detected Maternal CNV detected causing the false positive result

Lower the risk of false positive results determined by maternal CNVs Array-CGH Maternal blood PrenatalSafe Karyo cfdna