文献_2013-Evaluation of targeted next-generation sequencing–based preimplantation genetic diagnosis of monogenic disease.pdfVIP

ORIGINAL ARTICLE: GENETICS Evaluation of targeted next-generation sequencing–based preimplantation genetic diagnosis of monogenic disease Nathan R. Treff, Ph.D.,a,b,c Anastasia Fedick, B.S.,a,b Xin Tao, M.S.,a Batsal Devkota, Ph.D.,a Deanne Taylor, Ph.D.,a,c and Richard T. Scott Jr., M.D.a,c a Reproductive Medicine Associates of New Jersey, Morristown, New Jersey; b Molecular Genetics, Microbiology and Immunology, and c Obstetrics, Gynecology, and Reproductive Sciences, University of Medicine and Dentistry of New Jersey–Robert Wood Johnson Medical School, New Brunswick, New Jersey Objective: To investigate the applicability of next-generation sequencing (NGS) to preimplantation genetic diagnosis (PGD); to evaluate semiconductor-based NGS for genetic analysis of human embryos. Design: Blinded. Setting: Academic center for reproductive medicine. Patient(s): Six couples at risk of transmitting single gene disorders to their offspring. Intervention(s): None. Main Outcome Measure(s): Embryonic genotype consistency of NGS with two independent conventional methods of PGD. Result(s): NGS provided 100% equivalent PGD diagnoses of compound point mutations and small deletions and insertions compared with both reference laboratory– and internally developed quantitative polymerase chain reaction (qPCR)–based analyses. Furthermore, NGS single-gene disorder screening could be performed in parallel with qPCR-based comprehensive chromosome screening. Conclusion(s): NGS can provide blastocyst PGD results with a high level of consistency with established methodologies. This study and its design could serve as a model for further development of this important and emerging tech- nology. (Fertil Steril 2013; -:-– -. 2013 by American Society for Reproductive Medicine.) Use your smartphone Key Words: Next-generation sequencing, preimpla