The landscape for prenatal diagnostics varies widely across Europe, but in all cases there are dramatic shifts in the range of technology that is available, both for invasively-obtained samples and for non-invasive testing. Cytogenetic arrays and next-gen
sequencing are becoming much more common, but there are numerous key issues beyond just technology. Issues related to access, test interpretation, genetic counseling and implementation of newer tests will all have an impact on changes in the
Prenatal Diagnostics landscape in Europe.
Monday, 13 April
08:00 – 12:00 Preimplantation Genetic Diagnostics
In Vitro Fertilization (IVF) offers the opportunity for a range of genetic diagnostics to be carried out prior to the selection of fertilized eggs for implantation. In some cases a major reason for choosing the IVF route is because of the risk of
serious recessive genetic disorders, which can be detected by targeted screening. Screening and de-selection of eggs with aneuploidies is also a way of improving the success rate of IVF. Experience with arrays and with next-generation sequencing
will be discussed, as will recommendations for increasing the occurrence of singleton births.
- Joyce Harper, FRCPath, Genetics, Embryology and IVF Group, Institute for Womens Health, University College London, United Kingdom
- Jorish Vermeesch, Ph.D., Department of Human Genetics, Catholic University of Leuvan, Belgium
- Karsten R. Held, M.D., Medical Director, Reprogenetics Germany GmbH, Germany
- Francesco Fiorentino, Ph.D., Chief Executive Officer, GENOMA Group, Italy
- Tony Gordon, Ph.D., Laboratory Director (UK) and Managing Director (USA), Genesis Genetics
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* Separate registration required for short courses.
12:00 – 13:00 Registration
13:00 Chairperson’s Opening Remarks
13:05 Current Status, Issues and Challenges for Molecular Analysis of Invasively-Obtained Samples
Marta Rodriguez de Alba, Ph.D., Genetics, Fundacion Jimenez
The uptake of different types of molecular analysis for invasively obtained prenatal samples has varied considerably depending on technology and location. QF-PCR rapidly replaced FISH analysis for rapid aneuploidy diagnosis. The transition to Chromosomal
Microarray Analysis (CMA) has been slower, but recently has become much more widely accepted. Differences in the incorporation and reimbursement of CMA across countries in Europe will be presented. Some of the challenges for CMA, and approaches
for addressing these challenges, will also be discussed.
13:35 Prenatal Diagnosis Using Software-Targeted Array CGH: Robust, Rapid and Unequivocal
Joo Wook Ahn, Ph.D., Guy’s and St. Thomas’ NHS Foundation Trust
Prenatal testing of invasively-obtained samples should demonstrate robust performance and rapid turn-around times and should be affordable within a state-funded health service. Known pathogenic CNVs should be captured, and, ideally, equivocal results
should be avoided to minimize parental anxiety and delays in reporting. We have implemented a prenatal array CGH service using software-targeting, designed to detect CNVs of greater than 3Mb, and imbalance in regions of known pathogenicity. All
our results are therefore unequivocal and can be reported immediately. As new regions of pathogenicity are published, these can be added to the software-targeting, as can any loci relevant to specific ultrasound anomalies. This service started
in March 2012; the results of over 700 samples will be presented and discussed.
14:05 Confirmatory Invasive Testing after a Positive NIPT Result: Chorionic Villus Sampling or Amniocentesis?
Diane Van Opstal, Ph.D., Clinical Genetics, Erasmus Medical Center, The Netherlands
Non-invasive prenatal testing (NIPT) for fetal trisomy detection already revealed that there is a small chance of a false positive and false negative result. This is partly due to the fact that the fetal DNA present in the cell-free maternal plasma
fraction is derived from the cytotrophoblast of chorionic villi (CV). From cytogenetic studies in CV we know that the cytotrophoblast is not always representative for the fetus due to chromosomal mosaicism. Therefore, a positive NIPT should always
be confirmed with invasive testing in order to investigate the fetal karyotype. The fact that NIPT can be performed from the 10th week of gestation on makes CV sampling, routinely applied between 11-14 weeks of gestation, a more suitable technique
for confirmation studies than amniocentesis, mostly carried out after 15,5 gestational weeks. Based on our experience with cytogenetic investigations in CV, the choice for CV sampling or amniocentesis will highly depend on the chromosome aberration
involved. For trisomy 13, 18 and 21, we can recommend confirmation studies in CV, provided that these studies include the cytogenetic investigation of both the cytotrophoblast (for confirmation of the NIPT result) and the mesenchymal core (for
verification of the fetal karyotype). The protocol for other chromosome aberrations will be shown as well.
Utilization of a SNP Microarray for High-Resolution Prenatal Studies Over 15,000 Samples
Stuart Schwartz, Ph.D., Strategic Director, Cytogenetics, Laboratory Corporation
The findings of over 15,000 prenatal tests utilizing SNP microarray analysis will be reviewed. This data illustrates the utilization not only for ultrasound abnormalities, but also for patients referred for AMA and patients identified with chromosomal
anomalies that need better clarification. These studies have not only illustrated the importance of the detection of the gain or loss of chromosomal material, but also the importance of copy-neutral loss of heterozygosity (uniparental disomy
or identity by descent). It has also demonstrated the usefulness of examining the SNP alleles for detection of maternal cell contamination, twin-twin contamination, as well as mosaicism and whole genome homozygosity.
15:05 Refreshment Break
15:50 Advances in the Isolation and Analysis of Trophoblastic Cells for Non-Invasive Prenatal Diagnosis
Patrizia Paterlini-Brechot, Ph.D., Cellular and
Molecular Biology, University of Paris Descartes, France
Trophoblastic cells can be isolated non-invasively from blood and from the cervix at very early terms of pregnancy. The DNA of these fetal cells is not mixed with maternal DNA and can be used efficiently for prenatal, early and non-invasive prenatal
diagnosis. Results with consistent isolation and recovery of these cells, and subsequent genetic analysis of DNA from them will be presented. The advantages and remaining challenges for this approach will also be discussed.
16:20 Enrichment of Fetal Nucleated Red Blood Cells from Maternal Blood Using Novel Immunomagnetic Technology (CEPir) for Prenatal Diagnosis
Mahmoud Abuelhija, Ph.D., Research & Development, BioCEP Ltd., Israel
BioCEP has implemented a new method for Fetal Nucleated Red Blood Cells enrichment (fNRBC) from maternal blood, obtained at 8-14 weeks gestation. The new procedure is designed as an alternative to invasive prenatal testing, or to cell-free DNA
testing. This approach offers lower risk than invasive approaches, while offering better access to fetal DNA than is achieved with cell-free DNA. Challenges and opportunities for commercialization of this approach in the near future will be
16:50 ARCEDI - a Rapid Method for Isolation of Fetal Cells from Maternal Blood
Steen Kolvraa, M.D., Chief Scientific Officer, Arcedi Biotech ApS, Denmark
A method for isolation of fetal cells, presumably extravillous trophoblasts, will be presented. The method involves initial PFA fixation of full blood followed by immunomagnetic enrichment of fetal cells using endothelial markers, smearing
on slides and identification of fetal cells by ectodermal markers. The total method takes three days. In normal pregnancies we find around ten cells on 30 ml maternal blood.
17:20 Sponsored Presentation (Opportunity Available)
17:50 Close of Day One
TUESDAY, 14 April
8:00 Registration and Morning Coffee
9:00 Chairperson’s Remarks
Brigitte Faas, Ph.D., Human Genetics, Radboud University, The Netherlands
9:05 KEYNOTE PRESENTATION:
New Developments in Plasma DNA Sequencing for Non-Invasive Prenatal Testing
Y. M. Dennis Lo, M.D., Ph.D., Chairman, Chemical Pathology, Director, Li Ka Shing Institute of
Health Sciences, The Chinese University of Hong Kong, China
The current generation of noninvasive prenatal tests for fetal aneuploidies is based on the counting of DNA molecules in maternal plasma. We have developed a new approach based on the measurement of molecular size, a method that
exploits the fact that fetal DNA in maternal plasma is shorter than its maternal counterpart. Other exciting recent developments include the elucidation of the fetal methylome and transcriptome from maternal plasma. Such developments
have expanded the spectrum of research and diagnostic applications of noninvasive prenatal testing.
9:35 Challenges Associated with the Implementation of Whole Genome NIPT
Brigitte Faas, Ph.D., Human Genetics, Radboud University, The Netherlands
The implementation of whole genome NIPT goes along with several challenges, either from a political, counseling or technical point-of-view. In this presentation these challenges will be illustrated by examples and results from the
NIPT implementation study in the Netherlands and the Radboud University Nijmegen laboratory.
10:05 Implementing Cell-Free DNA Prenatal Testing into Clinical Practice in a Public Health Service
Suzanne Drury, Ph.D., Post Doctoral Scientist, NE Thames Regional Genetics Service, Great Ormond Street Hospital for Children, NHS Foundation Trust, United Kingdom
Implementing prenatal diagnosis based on cell free fetal DNA into public sector maternity care requires evaluation of laboratory performance, clinical utility and validity, health economics as well as patient and health professional
opinions. In the UK NIPD for selected monogenic disorders is now part of standard care, and the costs, benefits, uptake and overall impact of NIPT for aneuploidy in the NHS is being assessed.
Challenges of Developing a Regulated in vitro Product for NIPT
Stephen Little, Ph.D., CEO, Premaitha Health
Understanding the technical hurdles for launching a quality standard prenatal screening test to given consistency, reliability and robustness in a distrubuted environment. Importance of QA, QC, verification and validation to ensure
rapid and reliable uptake.
NIPT Performance in a Large General Population and Comparison Between High-Risk and Low-Risk Pregnancies
Grover Cong Yu, Ph.D., CEO, BGI-Europe & Africa, BGI Diagnostics
NIPT has been widely used to screen for T21, T18, and T13 in the past few years, yet large clinical data is still absent and concerns have been raised about the performance in large-scale clinical practice. Additionally, validation
of NIPT performance in low-risk population is still incomplete. To provide the large clinical data and evaluate the NIPT performance as an audit assessment, we initiated a multi-center prospective study to collect data from
the general population in Mainland China from January 1, 2012 to August 31, 2013. To assess the NIPT performance, sensitivity and specificity were calculated with the NIPT positive cases confirmed by karyotyping results, and
NIPT negative results confirmed by neonatal physical examination as well as telephone interview at one month after delivery. An insurance program was also used to encourage the report of NIPT false positve and false negative
results. Importantly, the study population was divided into high-risk and low-risk groups based on the maternal age, previous Down screening results, ultrasound finding, and pregnancy history. The NIPT performance in the high-risk
and low-risk populations was then compared.
11:05 Coffee Break in the Exhibit Hall with Poster Viewing
11:45 Sequencing of Cell-Free DNA as a Marker of Risk for Common Aneuploidies, Pre-Term Labor and Preeclampsia
Leona Poon, MRCOG, M.D., Consultant in Fetal Medicine and Obstetrics, Division
of Women’s Health, King’s College London, United Kingdom
Clinical implementation of cell-free DNA testing can either be as routine general screening or as contingent screening, based on other results, such as the first trimester combined test. A comparison of these two approaches,
and considerations for interpretation of cell-free DNA results, will be presented. We have also examined the potential for making use of cell-free DNA screening to detect increased risk of preeclampsia or spontaneous preterm
labor. Early results and challenges with both of these testing approaches will also be discussed.
12:15 Comparison of Genetic Signatures to Identify Prognostic Markers for the Risk of Pre-Term Birth
Joe Vockley, Ph.D., Chief Scientific Officer, Inova Translational Medical
Institute, Inova Fairfax Medical Center, Pediatrics, Virginia Commonwealth University School of Medicine, United States
The Inova Translational Medicine Institute has utilized whole genome sequences and other genomic data from thousands of families to develop a model for predicting the risk of pre-term birth. Data are generated from mother/father/baby
trios from a pre-term cohort and a full-term cohort to include families from over 100 countries. RNA markers were found in the peripheral blood of mothers that correlate to pre-term birth but not to common causes of pre-term
birth such as premature rupture of membranes and pre-eclampsia. These markers may be critical to the development of a tool for the prediction of preterm birth.
Luncheon Presentation: Advances in Next-Generation Sequencing for Non-Invasive Prenatal Testing
Alex Helm, MBA, Senior Product Manager, Illumina, Inc.
13:15 Session Break
14:15 Chairperson’s Remarks
14:20 Recovering High Fetal Fractions during Sample Preparation for NIPT: The Potential Role of Point-of-Care Technologies
Maiwenn Kersandy-Kerhoas, Ph.D.,
School of Engineering and Physical Sciences, Hariot Watt University, United Kingdom
Low fetal fraction of cell-free circulating DNA in the maternal circulation is the primary limitation met by most current prenatal molecular diagnostic technologies and results in sample rejection or lower test reliability.
To circumvent this limitation and preserve high fetal fraction, strict sample collection and preparation guidelines have been advocated by clinicians and analysts. Commercial cell-preservation solutions have entered the
market, however this area may benefit from a paradigm shift and high fetal fraction preservation and enrichment may lie in on-site sample preparation technologies. A comparative study revealing a 4-fold enrichment of fetal
fractions after on-chip maternal blood plasma extraction compared to conventional centrifugation will be presented.
14:50 Clinical Experience Using Targeted Microarrays for NIPT
Tuba Gunel, Ph.D., Molecular Biology and Genetics, Istanbul University, Turkey
Analysis of maternal blood plasma cell-free fetal (cff) DNA using array-CGH allows for detection of whole chromosome differences between test and reference DNA. Such arrays are commonly used to analyze amniotic samples and
pre-implantation embryos. We hypothesized that extraction, fluorescent labeling, hybridization, and analysis of cffDNA could be used to simultaneously screen for aneuploidy across every chromosome. This technology may also
aid the identification of minor genetic aberrations, such as micro-deletions and micro-duplications, which could enhance prenatal genetic diagnostics. NIPT using microarrays provided faster and more accurate cell-free DNA
(cfDNA) measurements than sequencing.
15:20 ANGELAB, Developing Lab-on-a-Chip-Based Systems for Epigenetic Non-Invasive Prenatal Diagnosis
Jesus M. Ruano-Lopez, Ph.D., IK4-Ikerlan, Spain
The European Project called ANGELAB, which is developing a family of In Vitro Diagnostic Systems, will be presented. Each LabonaChip runs fetal DNA sample preparation in combination with real-time or Digital PCR. The methodology
and system architecture used to transfer epigenetic biological protocols into modules, then into LabonaChip, and then into integrated systems will be described. The strategy, challenges and progress made over the first
two years of this project will also be covered.
15:50 Refreshment Break in the Exhibit Hall with Poster Viewing
16:30 PANEL DISCUSSION: NIPT Providers
Phillips Kuhl, President, Cambridge Healthtech Institute, United States
Maximilian Schmid, M.D., Associate
Director, Medical Affairs, Ariosa Diagnostics,United States
Alex Helm, Product Manager, lllumina Inc.,United States
Michael Lutz, Ph.D., Chief Executive Officer, LifeCodexx AG,
Matthew Hill, Ph.D., Vice President, Research and Development,
Natera, United States
John Anson, Ph.D., Executive Vice President, Research & Development,
Oxford Gene Technology, United Kingdom
Stephen Little, Chief Executive Officer, Premaitha Health,
Daniel Grosu, MD, Vice President, Chief Medical Officer, Research
& Development, Sequenom, United States
18:00 Welcome Reception in the Exhibit Hall with Poster Viewing
19:00 Close of Day Two
WEDNESDAY, 15 April
8:00 Registration and Morning Coffee
8:40 Chairperson’s Remarks
8:45 NIPD for Monogenic Disorders without Using Next-Generation Sequencing
Jessica van den Oever, MSc,
Department of Clinical Genetics, Laboratory for Diagnostic Genome Analysis (LDGA); Department of Clinical Genetics, Leiden University Medical Center
Next generation sequencing (NGS) is the main method used for non-invasive prenatal testing and diagnosis. However this method is still quite costly and may not be accessible to all laboratories. Moreover, although
the approach is very successful for detection of fetal aneuploidies it is also has its limitations. Noninvasive detection of fetal repeat expansions or paternally inherited mutations in certain regions of the
genome is currently difficult, if not impossible, by using NGS. A sensitive, fast and low cost alternative for NGS-based targeted mutation scanning will be presented, as well as a validation study for detection
of polymorphic paternally inherited CAG repeats for fetuses at risk of Huntington Disease.
9:15 Development of Quantitative PCR for NIPD of Single Gene Disorders
Claire Guissart, Ph.D., Laboratory of Genetic
Medicine, University of Montpellier, France
In recent years, non-invasive prenatal diagnosis (NIPD) has found new applications in monogenic disease diagnostics for paternally inherited mutations. This presentation discusses the application of a qPCR-based
mutant enrichment technique for NIPD of single gene disorder using Cystic Fibrosis as a clinical application model. In this respect, we have also repurposed a commercial mini-STR kit — frequently used
in forensic STR analysis — as an external quality control to confirm the presence of cell-free fetal DNA.
Experience with over 5000 NIPT Samples: Overcoming Technical and Biological Challenges
Joris R. Vermeesch, Ph.D., Professor, Molecular
Cytogenetics and Genome Research, University of Leuven
NIPT for fetal aneuploidy detection is increasingly being offered in the clinical setting. We present an analysis pipeline that enables the differentiation of fetal trisomies from local maternal CNVs and the detection
of non-classical aneuploidies as well as segmental imbalances. We present the results from its clinical application on over 4000 prospective pregnancies. Interestingly, our analysis pipeline also identified
a pregnant woman with early-stage nodular sclerosis Hodgkin lymphoma.
10:15 Coffee Break in the Exhibit Hall with Poster Viewing
10:45 The Diagnostic Accuracy of Non-Invasive Fetal RHD Typing
Florentine Thurik, M.D., Experimental Immunohematology, Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, The Netherlands
In the Netherlands non-invasive fetal RHD screening was implemented in July 2011. The evaluation of this screening program shows the high diagnostic accuracy of this approach; only 9 false negative results on >
26.000 serologically confirmed tests. Since the fetal RHD screening is based on a quantitative RHD PCR, we now have access to a huge data set on quantitative fetal DNA concentrations, showing the large biological
variability. The causes of false positivity and negativity will be discussed, and the lessons to be learned from this analysis for other non-invasive tests.
11:15 PANEL DISCUSSION: Predicting the Landscape for Prenatal Molecular Diagnostics in Europe
The current landscape for prenatal molecular diagnostics varies across Europe from one country to another. How might changes such as arraybased testing have an impact on invasively-obtained and possibly noninvasively
obtained samples? How soon might the isolation of fetal cells from maternal blood become commercially viable, and what would the implications of this shift be for NIPT? Are kit-based tests for NIPT more
likely to have an impact on NIPT in the short run? What other factors are likely to be important for changes in the clinical implementation of prenatal molecular diagnostics?
Marta Rodriguez de
Alba, Ph.D., Genetics, Fundacion Jimenez Diaz, Spain
Paterlini-Brechot, Ph.D., Cellular and Molecular Biology, University of Paris Descartes, France
Brigitte Faas, Ph.D.,
Human Genetics, Radboud University, The Netherlands
11:45 Close of Conference