title: Clinical outcome of preimplantation genetic diagnosis and screening using next generation sequencing.
abstract: BACKGROUND
  Next generation sequencing (NGS) is now being used for detecting chromosomal abnormalities in blastocyst trophectoderm (TE) cells from in vitro fertilized embryos. However, few data are available regarding the clinical outcome, which provides vital reference for further application of the methodology. Here, we present a clinical evaluation of NGS-based preimplantation genetic diagnosis/screening (PGD/PGS) compared with single nucleotide polymorphism (SNP) array-based PGD/PGS as a control.
  
  
  RESULTS
  A total of 395 couples participated. They were carriers of either translocation or inversion mutations, or were patients with recurrent miscarriage and/or advanced maternal age. A total of 1,512 blastocysts were biopsied on D5 after fertilization, with 1,058 blastocysts set aside for SNP array testing and 454 blastocysts for NGS testing. In the NGS cycles group, the implantation, clinical pregnancy and miscarriage rates were 52.6% (60/114), 61.3% (49/80) and 14.3% (7/49), respectively. In the SNP array cycles group, the implantation, clinical pregnancy and miscarriage rates were 47.6% (139/292), 56.7% (115/203) and 14.8% (17/115), respectively. The outcome measures of both the NGS and SNP array cycles were the same with insignificant differences. There were 150 blastocysts that underwent both NGS and SNP array analysis, of which seven blastocysts were found with inconsistent signals. All other signals obtained from NGS analysis were confirmed to be accurate by validation with qPCR. The relative copy number of mitochondrial DNA (mtDNA) for each blastocyst that underwent NGS testing was evaluated, and a significant difference was found between the copy number of mtDNA for the euploid and the chromosomally abnormal blastocysts. So far, out of 42 ongoing pregnancies, 24 babies were born in NGS cycles; all of these babies are healthy and free of any developmental problems.
  
  
  CONCLUSIONS
  This study provides the first evaluation of the clinical outcomes of NGS-based pre-implantation genetic diagnosis/screening, and shows the reliability of this method in a clinical and array-based laboratory setting. NGS provides an accurate approach to detect embryonic imbalanced segmental rearrangements, to avoid the potential risks of false signals from SNP array in this study.
authors: Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha, China ; National Engineering and Research Center of Human Stem Cell, Changsha, China ; Reproductive & Genetic Hospital of CITIC Xiangya, Changsha, China.
authors: Yueqiu
authors: Y
authors: Tan
authors: BGI-Health, BGI-Shenzhen, Shenzhen, China ; Shenzhen Municipal Birth Defect Screening Project Lab, BGI-Shenzhen, Shenzhen, China ; Guangdong Enterprise Key Laboratory of Human Disease Genomics, BGI-Shenzhen, Shenzhen, China.
authors: Xuyang
authors: X
authors: Yin
authors: Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha, China ; Reproductive & Genetic Hospital of CITIC Xiangya, Changsha, China ; Key Laboratory of Stem Cell and Reproductive Engineering, Ministry of Health, Changsha, China.
authors: Shuoping
authors: S
authors: Zhang
authors: Shenzhen Municipal Birth Defect Screening Project Lab, BGI-Shenzhen, Shenzhen, China ; Guangdong Enterprise Key Laboratory of Human Disease Genomics, BGI-Shenzhen, Shenzhen, China ; Department of Biology, University of Copenhagen, Copenhagen, Denmark.
authors: Hui
authors: H
authors: Jiang
authors: Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha, China ; National Engineering and Research Center of Human Stem Cell, Changsha, China.
authors: Ke
authors: K
authors: Tan
authors: BGI-ShenZhen, ShenZhen, China.
authors: Jian
authors: J
authors: Li
authors: Reproductive & Genetic Hospital of CITIC Xiangya, Changsha, China.
authors: Bo
authors: B
authors: Xiong
authors: Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha, China ; Reproductive & Genetic Hospital of CITIC Xiangya, Changsha, China.
authors: Fei
authors: F
authors: Gong
authors: Shenzhen Municipal Birth Defect Screening Project Lab, BGI-Shenzhen, Shenzhen, China ; Guangdong Enterprise Key Laboratory of Human Disease Genomics, BGI-Shenzhen, Shenzhen, China.
authors: Chunlei
authors: C
authors: Zhang
authors: Shenzhen Municipal Birth Defect Screening Project Lab, BGI-Shenzhen, Shenzhen, China ; Guangdong Enterprise Key Laboratory of Human Disease Genomics, BGI-Shenzhen, Shenzhen, China ; School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, China.
authors: Xiaoyu
authors: X
authors: Pan
authors: Shenzhen Municipal Birth Defect Screening Project Lab, BGI-Shenzhen, Shenzhen, China ; Guangdong Enterprise Key Laboratory of Human Disease Genomics, BGI-Shenzhen, Shenzhen, China ; Section of Molecular Disease Biology, Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors: Fang
authors: F
authors: Chen
authors: Shenzhen Municipal Birth Defect Screening Project Lab, BGI-Shenzhen, Shenzhen, China ; Guangdong Enterprise Key Laboratory of Human Disease Genomics, BGI-Shenzhen, Shenzhen, China ; State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, China.
authors: Shengpei
authors: S
authors: Chen
authors: BGI-ShenZhen, ShenZhen, China.
authors: Chun
authors: C
authors: Gong
authors: Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha, China ; Reproductive & Genetic Hospital of CITIC Xiangya, Changsha, China.
authors: Changfu
authors: C
authors: Lu
authors: Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha, China ; Reproductive & Genetic Hospital of CITIC Xiangya, Changsha, China.
authors: Keli
authors: K
authors: Luo
authors: Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha, China ; Reproductive & Genetic Hospital of CITIC Xiangya, Changsha, China.
authors: Yifan
authors: Y
authors: Gu
authors: Guangdong Enterprise Key Laboratory of Human Disease Genomics, BGI-Shenzhen, Shenzhen, China.
authors: Xiuqing
authors: X
authors: Zhang
authors: BGI-Health, BGI-Shenzhen, Shenzhen, China ; Shenzhen Municipal Birth Defect Screening Project Lab, BGI-Shenzhen, Shenzhen, China.
authors: Wei
authors: W
authors: Wang
authors: BGI-ShenZhen, ShenZhen, China.
authors: Xun
authors: X
authors: Xu
authors: BGI-ShenZhen, ShenZhen, China ; Central Queensland University, Rockhampton, Queensland Australia.
authors: Gábor
authors: G
authors: Vajta
authors: BGI-ShenZhen, ShenZhen, China ; Department of Biomedicine, Aarhus University, Aarhus, Denmark.
authors: Lars
authors: L
authors: Bolund
authors: BGI-ShenZhen, ShenZhen, China ; Prince Aljawhra Center of Excellence in Research of Hereditary Disorders, King Abdulaziz University, Jeddah, Saudi Arabia ; James D Watson Institute of Genome Science, Hangzhou, China.
authors: Huanming
authors: H
authors: Yang
authors: Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha, China ; National Engineering and Research Center of Human Stem Cell, Changsha, China ; Reproductive & Genetic Hospital of CITIC Xiangya, Changsha, China ; Key Laboratory of Stem Cell and Reproductive Engineering, Ministry of Health, Changsha, China.
authors: Guangxiu
authors: G
authors: Lu
authors: BGI-Health, BGI-Shenzhen, Shenzhen, China.
authors: Yutao
authors: Y
authors: Du
authors: Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha, China ; National Engineering and Research Center of Human Stem Cell, Changsha, China ; Reproductive & Genetic Hospital of CITIC Xiangya, Changsha, China ; Key Laboratory of Stem Cell and Reproductive Engineering, Ministry of Health, Changsha, China.
authors: Ge
authors: G
authors: Lin
keywords: Blastocyst; Clinical outcome; Cryopreserved embryo transfer; Next generation sequencing; Preimplantation genetic diagnosis/screening