title: Evaluation of PCR-based preimplantation genetic diagnosis applied to monogenic diseases: a collaborative ESHRE PGD consortium study.
abstract: Preimplantation genetic diagnosis (PGD) for monogenic disorders currently involves polymerase chain reaction (PCR)-based methods, which must be robust, sensitive and highly accurate, precluding misdiagnosis. Twelve adverse misdiagnoses reported to the ESHRE PGD-Consortium are likely an underestimate. This retrospective study, involving six PGD centres, assessed the validity of PCR-based PGD through reanalysis of untransferred embryos from monogenic-PGD cycles. Data were collected on the genotype concordance at PGD and follow-up from 940 untransferred embryos, including details on the parameters of PGD cycles: category of monogenic disease, embryo morphology, embryo biopsy and genotype assay strategy. To determine the validity of PCR-based PGD, the sensitivity (Se), specificity (Sp) and diagnostic accuracy were calculated. Stratified analyses were also conducted to assess the influence of the parameters above on the validity of PCR-based PGD. The analysis of overall data showed that 93.7% of embryos had been correctly classified at the time of PGD, with Se of 99.2% and Sp of 80.9%. The stratified analyses found that diagnostic accuracy is statistically significantly higher when PGD is performed on two cells versus one cell (P=0.001). Se was significantly higher when multiplex protocols versus singleplex protocols were applied (P=0.005), as well as for PGD applied on cells from good compared with poor morphology embryos (P=0.032). Morphology, however, did not affect diagnostic accuracy. Multiplex PCR-based methods on one cell, are as robust as those on two cells regarding false negative rate, which is the most important criteria for clinical PGD applications. Overall, this study demonstrates the validity, robustness and high diagnostic value of PCR-based PGD.
authors.affiliation: Departments of Clinical Genetics and School for Oncology and Developmental Biology, GROW, Maastricht University Medical Centre, Maastricht, The Netherlands.
authors.forename: Jos
authors.initials: J
authors.lastname: Dreesen
authors.affiliation: Laboratory of Medical Genetics, University of Athens, St Sophia's Children's Hospital, Athens, Greece.
authors.forename: Aspasia
authors.initials: A
authors.lastname: Destouni
authors.affiliation: The Stavros Niarchos Foundation-Collaborative Center for Clinical Epidemiology and Outcomes Research (CLEO), First and Second University Department of Pediatrics, National and Kapodistrian University of Athens, School of Medicine, Athens, Greece.
authors.forename: Georgia
authors.initials: G
authors.lastname: Kourlaba
authors.affiliation: Fertility Clinic, Department of Obstetrics and Gynaecology, Aarhus University Hospital, Skejby DK-8200 Aarhus N, Denmark.
authors.forename: Birte
authors.initials: B
authors.lastname: Degn
authors.affiliation: Fertility Clinic, Department of Obstetrics and Gynaecology, Aarhus University Hospital, Skejby DK-8200 Aarhus N, Denmark.
authors.forename: Wulf Christensen
authors.initials: WC
authors.lastname: Mette
authors.affiliation: Department of Genetics, Faculty of Medicine, University of Porto, Porto, Portugal.
authors.forename: Filipa
authors.initials: F
authors.lastname: Carvalho
authors.affiliation: Université de Strasbourg FRANCE & HUS - Service de la Biologie de la Reproduction, CMCO, 19, Rue Louis Pasteur, BP120, Schiltigheim, France.
authors.forename: Celine
authors.initials: C
authors.lastname: Moutou
authors.affiliation: UCL Centre for PG&D, Institute for Women's Health, London, UK.
authors.forename: Sioban
authors.initials: S
authors.lastname: Sengupta
authors.affiliation: UCL Centre for PG&D, Institute for Women's Health, London, UK.
authors.forename: Seema
authors.initials: S
authors.lastname: Dhanjal
authors.affiliation: Guy's & St Thomas' Centre for Preimplantation Genetic Diagnosis and Genetics, Guy's & St Thomas' NHS Foundation Trust, London, UK.
authors.forename: Pamela
authors.initials: P
authors.lastname: Renwick
authors.affiliation: Embryogenesis Assisted Conception Unit, Athens, Greece.
authors.forename: Steven
authors.initials: S
authors.lastname: Davies
authors.affiliation: Laboratory of Medical Genetics, University of Athens, St Sophia's Children's Hospital, Athens, Greece.
authors.forename: Emmanouel
authors.initials: E
authors.lastname: Kanavakis
authors.affiliation: Reprogenetics, 3 Regent Street, Suite 301, Livingston, NJ, USA.
authors.forename: Gary
authors.initials: G
authors.lastname: Harton
authors.affiliation: Laboratory of Medical Genetics, University of Athens, St Sophia's Children's Hospital, Athens, Greece.
authors.forename: Joanne
authors.initials: J
authors.lastname: Traeger-Synodinos
mesh_terms: D001706:Biopsy; D001757:Blastomeres / Q000378:metabolism; D005260:Female; D030342:Genetic Diseases, Inborn / Q000175:diagnosis* / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D019836:Preimplantation Diagnosis / Q000379:methods; D015203:Reproducibility of Results; D012189:Retrospective Studies; D012307:Risk Factors