Non-invasive prenatal testing (NIPT): Europe’s first multicenter post-market clinical follow-up study validating the quality in clinical routine



Non-invasive prenatal tests (NIPT) for the determination of fetal aneuploidies from maternal blood are firmly established in clinical routine. For the first time, the accuracy of an NIPT for the determination of trisomies 21, 18 and 13 in singleton pregnancies was assessed by means of a prospective German-wide multicenter post-market clinical follow-up study, to reliably evaluate the quality in clinical routine.


The study covered the indications for testing, the test results, the rate of invasive diagnostics and the pregnancy outcome. 2232 cases were tested for trisomy 21. Of these, 1946 cases were additionally examined for trisomy 18 and 13.


Sensitivity and specificity for trisomy 21 (43/43) and for trisomy 13 (2/2) were 100%, for trisomy 18 the sensitivity was 80% (4/5) with a specificity of 99.8%. Three false-positive results for trisomy 18 were observed (FPR 0.15%). The no-call rate was 0.5%. In this subgroup, 27.3% (3/11) aneuploidies were diagnosed. The rate of invasive procedures was 2.6%.


NIPT provides a very high quality for the fetal trisomies 21, 13 and 18 in clinical routine. The results support the recommendation that NIPT should be offered after genetic counseling and only in conjunction with a qualified ultrasound examination.

This is a preview of subscription content, log in to check access.

Fig. 1


  1. 1.

    Holzgreve W, Hahn S, Zhong XY et al (2007) Fetal genetic communication between fetus and mother: short- and long-term consequences. Am J Obstet Gynecol 196:372–381

    Article  PubMed  Google Scholar 

  2. 2.

    Mangeold-Bauer G, Holzgreve W, Geipel A (2016) Nichtinvasive Pränataldiagnostik—für jede Schwangere die Methode der Wahl? Gynäkologische Praxis 40:577–585

    Google Scholar 

  3. 3.

    Lo YMD, Corbetta N, Chamberlain PF et al (1997) Presence of fetal DNA in maternal plasma and serum. Lancet 350:485–487

    CAS  Article  PubMed  Google Scholar 

  4. 4.

    Zhong XY, Hahn S, Holzgreve W (2001) Prenatal identification of fetal genetic traits. Lancet 357:310–311

    CAS  Article  PubMed  Google Scholar 

  5. 5.

    Li Y, Hahn S, Holzgreve W (2006) Recent development in the detection of fetal single gene differences in maternal plasma and the role of size fractination. Ann N Y Acad Sci 1092:285–292

    CAS  Article  PubMed  Google Scholar 

  6. 6.

    Futch T, Spinosa J, Bhatt S, de Feo E, Rava RP, Sehnert AJ (2013) Initial clinical laboratory experience in noninvasive prenatal testing for fetal aneuploidy from maternal plasma DNA samples. Prenat Diagn 33:569–574

    Article  PubMed  PubMed Central  Google Scholar 

  7. 7.

    Palomaki GE, Kloza EM, Lambert-Messerlian GM et al (2011) DNA sequencing of maternal plasma to detect Down syndrome: an international clinical validation study. Genet Med 13:913–920

    CAS  Article  PubMed  Google Scholar 

  8. 8.

    Stumm M, Entezami M, Trunk N et al (2012) Noninvasive prenatal detection of chromosomal aneuploidies using different next generation sequencing strategies and algorithms. Prenat Diagn 32:569–577

    Article  PubMed  Google Scholar 

  9. 9.

    Norton ME, Brar H, Weiss J, Karimi A et al (2012) Non-Invasive Chromosomal Evaluation (NICE) Study: results of a multicenter prospective cohort study for detection of fetal trisomy 21 and trisomy 18. Am J Obstet Gynecol 207:137.e1–137.e8

    Article  Google Scholar 

  10. 10.

    Dar P, Curnow KJ, Gross SJ et al (2014) Clinical experience and follow-up with large scale single-nucleotide polymorphism-based noninvasive prenatal aneuploidy testing. Am J Obstet Gynecol 211:527.e1–527.e17

    Article  Google Scholar 

  11. 11.

    Cuckle H, Benn P, Pergament E (2015) Cell-free DNA screening for fetal aneuploidy as a clinical service. Clin Biochem 48:932–941

    CAS  Article  PubMed  Google Scholar 

  12. 12.

    Wagner AJ, Mitchell ME, Tomita-Mitchell A (2014) Use of cell-free fetal DNA in maternal plasma for noninvasive prenatal screening. Clin Perinatol 41:957–966

    Article  PubMed  Google Scholar 

  13. 13.

    Akolekar R, Beta J, Picciarelli G, Ogilvie C, D’Antonio F (2015) Procedure-related risk of miscarriage following amniocentesis and chorionic villus sampling: a systematic review and meta-analysis. Ultrasound Obstet Gynecol 45:16–26

    CAS  Article  PubMed  Google Scholar 

  14. 14.

    Lefkowitz RB, Tynan JA, Liu T et al (2016) Clinical validation of a noninvasive prenatal test for genomewide detection of fetal copy number variants. Am J Obstet Gynecol 215(2):227.e1–227.e16

    Article  Google Scholar 

  15. 15.

    Gil MM, Accurti V, Santacruz B, Plana MN, Nicolaides KH (2017) Analysis of cell-free DNA in maternal blood in screening for fetal aneuploidies: updated meta-analysis. Ultrasound Obstet Gynecol. doi:10.1002/uog.17484

    PubMed  Google Scholar 

  16. 16.

    Stumm M, Entezami M, Haug K et al (2014) Diagnostic accuracy of random massively parallel sequencing for non-invasive prenatal detection of common autosomal aneuploidies: a collaborative study in Europe. Prenat Diagn 34:185–191

    Article  PubMed  Google Scholar 

  17. 17.

    Verweij EJ, Jacobsson B, van Scheltema PA et al (2013) European non-invasive trisomy evaluation (EU-NITE) study: a multicenter prospective cohort study for non-invasive fetal trisomy 21 testing. Prenat Diagn 33:996–1000

    CAS  PubMed  Google Scholar 

  18. 18.

    Vora NL, Robinson S, Hardisty EE, Stamilio DM (2016) The utility of a prerequisite ultrasound at 10–14 weeks in cell free DNA fetal aneuploidy screening. Ultrasound Obstet Gynecol. doi:10.1002/uog.15995

    Google Scholar 

  19. 19.

    Larion S, Warsof SL, Romary L, Mlynarczyk M, Peleg D, Abuhamad AZ (2014) Uptake of noninvasive prenatal testing at a large academic referral center. Am J Obstet Gynecol 211:651.e1–651.e7

    Article  Google Scholar 

  20. 20.

    Salomon LJ, Alfirevic Z, Audibert F, Kagan KO, Paladini D, Yeo G, Raine-Fenning N (2017) ISUOG updated consensus statement on the impact of cfDNA aneuploidy testing on screening policies and prenatal ultrasound practice. Ultrasound Obstet Gynecol 49:815–816

    CAS  Article  PubMed  Google Scholar 

  21. 21.

    Nicolaides KH, Syngelaki A, Ashoor G, Birdir C, Touzet G (2012) Noninvasive prenatal testing for fetal trisomies in a routinely screened first-trimester population. Am J Obstet Gynecol 207:374.e1–374.e6

    Article  Google Scholar 

  22. 22.

    Bianchi DW, Parker RL, Wentworth J et al (2014) CARE Study Group. DNA sequencing versus standard prenatal aneuploidy screening. N Engl J Med 370:799–808

    CAS  Article  PubMed  Google Scholar 

  23. 23.

    Quezada MS, Gil MM, Francisco C, Oro`sz G, Nicolaides KH (2015) Screening for trisomies 21, 18 and 13 by cell-free DNA analysis of maternal blood at 10–11 weeks’ gestation and the combined test at 11–13 weeks. Ultrasound Obstet Gynecol 45:36–41

    CAS  Article  PubMed  Google Scholar 

  24. 24.

    Norton ME, Jacobsson B, Swamy GK et al (2015) Cell-free DNA analysis for noninvasive examination of trisomy. N Engl J Med 372:1589–1597

    CAS  Article  PubMed  Google Scholar 

  25. 25.

    Benn P, Borrell A, Chiu RW et al (2015) Position statement from the Chromosome Abnormality Screening Committee on behalf of the Board of the International Society for Prenatal Diagnosis. Prenat Diagn 35:725–734

    Article  PubMed  Google Scholar 

  26. 26.

    Gregg AR, Skotko BG, Benkendorf JL et al (2016) Noninvasive prenatal screening for fetal aneuploidy, 2016 update: a position statement of the American College of Medical Genetics and Genomics. Genet Med 18:1056–1065

    CAS  Article  PubMed  Google Scholar 

  27. 27.

    Schmid M, Klaritsch P, Arzt W et al (2015) Cell-Free DNA testing for fetal chromosomal anomalies in clinical practice: Austrian-German-Swiss recommendations for non-invasive prenatal tests (NIPT). Ultraschall Med 36:507–510

    CAS  Article  PubMed  Google Scholar 

  28. 28.

    Yaron Y (2016) The implications of non-invasive prenatal testing failures: a review of an under-discussed phenomenon. Prenat Diagn 36:391–396

    CAS  Article  PubMed  Google Scholar 

  29. 29.

    Manegold-Brauer G, Berg C, Flöck A, Rüland A, Gembruch U, Geipel A (2015) Uptake of non-invasive prenatal testing (NIPT) and impact on invasive procedures in a tertiary referral center. Arch Gynecol Obstet 292:543–548

    Article  PubMed  Google Scholar 

  30. 30.

    Petersen OB, Vogel I, Ekelund C, Hyett J, Tabor A (2014) Danish Fetal Medicine Study Group; Danish Clinical Genetics Study Group. Potential diagnostic consequences of applying non-invasive prenatal testing: population-based study from a country with existing first-trimester screening. Ultrasound Obstet Gynecol 43:265–271

    CAS  Article  PubMed  Google Scholar 

  31. 31.

    Revello R, Sarno L, Ispas A, Akolekar R, Nicolaides KH (2016) Screening for trisomies by cell-free DNA testing of maternal blood: consequences of a failed result. Ultrasound Obstet Gynecol 47:698–704

    CAS  Article  PubMed  Google Scholar 

  32. 32.

    Khalil A, Mahmoodian N, Kulkarni A et al (2015) Estimation of detection rates of aneuploidy in high-risk pregnancy using an approach based on nuchal translucency and non-invasive prenatal testing: a cohort study. Fetal Diagn Ther 38:254–261

    Article  PubMed  Google Scholar 

  33. 33.

    Warsof SL, Larion S, Abuhamad AZ (2015) Overview of the impact of noninvasive prenatal testing on diagnostic procedures. Prenat Diagn 35:972–979

    Article  PubMed  Google Scholar 

  34. 34.

    Kagan KO, Eiben B, Kozlowski P (2014) Combined first trimester screening and cell-free fetal DNA—“next generation screening”. Ultraschall Med 35:229–236

    CAS  Article  PubMed  Google Scholar 

  35. 35.

    Kagan KO, Sonek J, Wagner P, Hoopmann M (2017) Principles of first trimester screening in the age of non-invasive prenatal diagnosis: screening for chromosomal abnormalities. Arch Gynecol Obstet. doi:10.1007/s00404-017-4459-9

    Google Scholar 

Download references


First and foremost, we would like to thank the pregnant women who contributed to this study. Furthermore, we would like to thank all participating physicians for recruiting of the patients and the excellent collaboration.

Author information




AF contributed to protocol/project development, data collection or management, data analysis, manuscript writing/editing; NCT, data collection or management; AR, data collection or management; WH, protocol/project development, manuscript writing/editing; UG, manuscript writing/editing; AG, protocol/project development, manuscript writing/editing.

Corresponding author

Correspondence to Annegret Geipel.

Ethics declarations


This study was funded by the company LifeCodexx. University Medical School Bonn received 40.000€.

Conflict of interest

All authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in study involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Human/animal rights statement

This article does not contain any studies with animals performed by any of the authors.

Informed consent

Informed written consent was obtained from all individual participants included in the study.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Flöck, A., Tu, N., Rüland, A. et al. Non-invasive prenatal testing (NIPT): Europe’s first multicenter post-market clinical follow-up study validating the quality in clinical routine. Arch Gynecol Obstet 296, 923–928 (2017).

Download citation


  • NIPT
  • cfDNA
  • Next-generation sequencing
  • Screening for aneuploidies
  • Trisomy 21