NIPT NON – INVASIVE PRENATAL TEST GENDIA Antwerp, Belgium NIPT NON – INVASIVE PRENATAL TEST Testing of cff DNA (cell free fetal DNA) from maternal blood during pregnancy for trisomy 21, 18 and 13 www.DOWNsyndromeNIPT.info Frequency of fetal aneuploidies Aneuploidy Syndrome Frequency (live births) Trisomy 21 Down syndrome 1 in 700 Trisomy 18 Edwards syndrome 1 in 5,000 Trisomy 13 Patau syndrome 1 in 16,000 Trisomy 21, 18, 13 screening Trisomy 21 (Down syndrome) Trisomy 18 (Edwards syndrome) Trisomy 13 (Patau syndrome) Risk Down syndrome versus Maternal Age Age Frequency (live births) < 35 37 < 0.3 % 0.5 % 40 1% 50 10 % History of Down syndrome screening • 1980 : Amniocentesis (advanced maternal age) • 1990 : Triple screening (T21, T18 and T13) • 2000 : First trimester screening (T21, T18 and T13) • 2013 : Start of NIPT • 2015 : NIPT greatly replaces first trimester screening First trimester serum screening (FTS - combi test) Combi test : Risk calculated from : 1. Maternal age : the higher the age, the higher the risk of T21, T18, T13 2. Nuchal translucency (NT) : the higher the NT, the higher the risk of T21, T18, T13 3. Serum parameters PAPP-A and free B HCG If the combi risk is increased ( > 1/300 in Belgium or > 1/200 in The Netherlands) an invasive procedure (CVS / AC) or NIPT is recommended Classical Down syndrome screening NT (mm) PAPP-A (MoM) B-HCG (MoM) Normal : 2.0 Normal : 1.0 Normal : 1.0 T21 : 3.4 T21 : 0.5 T21 : 2.0 T18 : 5.5 T18 : 0.2 T18 : 0.2 T13 : 4.0 T13 : 0.3 T13 : 0.5 Nicolaides et al 2008 NIPT history • 1997 : Lo et al. :cff DNA in maternal circulation • 2001 : Fetal Rh(D) genotype • 2006 : Sexing fetus for : 1. X-linked genetic disorders 2. Sexing (China) • 2011 : Detection trisomy 21/18/13 becomes possible • 2012 : > 100.000 patients screened in China / USA • 2013 : GENDIA starts with NIPT in Belgium and the Netherlands • 2015 : GENDIA has completed 10.000 NIPT tests NIPT essentials 1. TEST : trisomy 21 (Down syndrome), trisomy 18 (Edwards syndrome), trisomy 13 (Patau syndrome). Also sex of the fetus is determined. 2. SAMPLE: Specific test kits provided by GENDIA 3. TIMING: > week 10 (preferentially week 11-12) 4. TURNAROUND TIME: < 2 weeks 5. RELIABILITY: 99% for trisomy 21 6. INDICATIONS: Although NIPT can be performed in every pregnancy, it is especially indicated: • If the triple test or first trimester screening indicates an increased risk • Advanced maternal age • Anxiety for invasive procedures 7. CONTRAINDICATIONS: NIPT is not the test of choice when there is : • Fetal anomalies on ultrasound • Severely elevated NT (nuchal translucency) with normal PAPP-A and free B HCG • A triplet pregnancy, vanished twin Cell Free Fetal DNA (cff DNA) in Maternal Blood NIPT cff DNA • < 1 % of total DNA in maternal circulation is fetal • 5-30 % of cell-free DNA in maternal circulation is fetal NIPT for trisomy 21 NIPT measures the ratio of chromosome 21 sequence versus control chromosome sequence to exclude trisomy 21 NIPT cffDNA cffDNA Nl T21 DNA Nl T21 Fetal Maternal • <1 of Importance of fetal fraction Fetal Fraction Expected ratio for Trisomy 4% 1.02 Maternal 10% 1.05 cfDNA 20% 1.10 40% 1.20 Fetal cfDNA Reference Chromosome Chromosome 21 Sensitivity of NIPT for T21, T18, T13 Sensitivity T21 : 99 % T18 : 98 % T13 : 90 % False-negatives If NIPT is normal, the residual risk for trisomy 21, 18, 13 < 1 / 10.000 Specitivity NIPT for T21, T18, T13 Specitivity T21 > 99 % T18 > 99 % T13 > 99 % False-positives If NIPT is abnormal, the risk that the fetus has no trisomy 21, 18, 13 is very small NO NIPT for sex aneuploidies • Phenotype for sex aneuploidies is highly variable • Mosaicism in the fetus is a problem • Mosaicism in the mother is a problem • NIPT for sex aneuploidies is less accurate NIPT Indications NIPT is the test of choice when there is : • Increased maternal age • Increased risk on Combination or triple test • Anxiety for invasive procedure (AC / CVS) NIPT Contra indications NIPT is NOT the test of choice when there is : • Fetal anomalies on ultrasound • A triplet pregnancy • Vanishing twin • Known genetic anomalies that cannot be diagnosed by NIPT NIPT Advantages versus combi test with AC / CVS • High sensitivity (few false-negatives) • High specificity (few false-positives) • More than T21 • Non-invasive : no fetal risk • CVS : Risk of miscarriage : 1-2 % • AC : Risk of miscarriage : 0.5 % NIPT versus FTS FTS NIPT False negatives > 20 % 2% False positives 95 % 2% NIPT versus classical screening Classical AC / CVS NIPT Safe Not safe Safe Not reliable Reliable Reliable Companies offering NIPT • ARIOSA (US) • VERINATA (US) • NATERA (US) • SEQUENOM (US) • BGI (China) • LIFE-CODEXX (Germany) Academic Hospitals offering NIPT None of the genetic centers in the academic centres in Belgium or the Netherlands currently offers a validated NIPT test with published results GENDIA offers the HARMONY test from ARIOSA • ARIOSA (Harmony) GENDIA offers the Harmony test from ARIOSA • Almost 400.000 Harmony tests performed • > 20.000 tests in clinical trials • > 20 scientific papers • Very low false-positive and false-negative rate • Low failure rate (0.6 %) • Fast turnaround NIPT results 1. Normal result : no specific follow up necessary, unless ultrasound examination of the fetus reveals anomalies 2. Test failure : in < 2 % pregnancies not enough fetal DNA : NIPT repeated at no extra cost. 3. Abnormal NIPT result : confirmation by amniocentesis or chorionic biopsy NIPT versus FTS in Belgium FTS NIPT Number screenings 100.000 100.000 Expected T21 200 (1/500) 200 (1/500) Detection rate 75 % 99 % T21 150 198 False-negatives 50 (25 %) 2 (1 %) False-positives 4990 (4.8 %) < 100 (0.03 %) Iatrogenic Miscarriage 50 1 NIPT results GENDIA • Total number NIPT : 10.000 • Total number of trisomies : 131 » T21 : 111 » T18 : 11 » T13 : 9 • 1 false-positives (T13) • 4 false-negatives (T21, T13, twice T18) • 2 inconclusive results (T18, T13) • No result : 0.7 % How offer NIPT to your patients ? 1. Refer to the consultation : Email : NIPT@GENDIA.net to ask for an appointment 2. Take blood yourself with our sample kit : Email : NIPT@GENDIA.net to ask for kits www.DOWNsyndromeNIPT.info www.DOWNsyndromeNIPT.info