The clinical application of preimplantation genetic diagnosis for the patient affected by congenital contractural arachnodactyly and spinal and bulbar muscular atrophy

Abstract

Purpose To investigate the usefulness of preimplantation genetic diagnosis (PGD) for the patient affected by congenital contractural arachnodactyly (CCA) and spinal and bulbar muscular atrophy (SBMA). Methods Multiple displacement amplification (MDA) was performed for whole genome amplification (WGA) of biopsied trophectoderm (TE) cells. Direct mutation detection by sequencing and next-generation sequencing (NGS)-based single nucleotide polymorphism (SNP) haplotyping were used for CCA diagnosis. Direct sequencing of the PCR products and sex determination by amplification of sexdetermining region Y (SRY) gene were used for SBMA diagnosis. After PGD, the unaffected blastocyst (B4) was transferred in the following frozen embryo transfer (FET). Results In this PGD cycle, sixteen MII oocytes were inseminated by ICSI with testicular spermatozoa. Four blastocysts (B4, B5, B10, B13) were utilized for TE cell biopsy on day 5 after ICSI. After PGD, B4 was unaffected by CCA and SBMA. B5 was affected by CCA and carried SBMA. B10 was unaffected by CCA and carried SBMA. B13 was affected by CCA and unaffected by SBMA. B4 was the only unaffected blastocyst and transferred into the uterus for the subsequent FET cycle. The accuracy of PGD was confirmed by amniocentesis at 21 weeks of gestation. A healthy boy weighing 2850 g was born by cesarean section at the 38th week of gestation. Conclusions PGD is a valid screening tool for patienst affected of CCA and SBMA to prevent transmission of these genetic diseases from parents to children.Keywords Congenital contractural arachnodactyly . Spinal and bulbar muscular atrophy . PGD . Frozen embryo transfer Capsule A description of a PGD cycle for the diagnosis of CCA and SBMA, resulting in the birth of a healthy boy.Linjun Chen and Zhenyu Diao contributed equally to this work.

The clinical application of single-sperm-based SNP haplotyping for PGD of osteogenesis imperfecta

“…Moreover, another advantage of haplotyping is the ability to identify meiotic recombination, which can impair the accuracy of PGD (Altarescu et al 2008). We previously diagnosed congenital contractural arachnodactyly via NGS-based SNP haplotyping and observed homologous recombination of maternal origin, but this effect did not impair the accuracy of PGD due to the more informative SNPs used in this case (Chen et al 2016). Therefore, haplotyping presents a number of advantages over the direct mutation detection by PCR, including the detection of ADO, chromosomal monosomy and recombination, to ensure the accuracy of clinical diagnosis.…”

“…The European Society of Human Reproduction and Embryology (ESHRE) PGD consortium reported that the misdiagnosis rate of the PCR-based method is 0.10% (13/12,790) (De Rycke et al 2017). Several polymorphic markers [short tandem repeats (STRs) or single-nucleotide polymorphisms (SNPs)] located upstream and downstream of the pathogenic genes associated with mutation sites have been used for linkage analysis to establish haplotypes for identifying ADO and to avoid misdiagnosis in PGD (Qubbaj et al 2011;Renwick et al 2006Renwick et al , 2010Chen et al 2016Chen et al , 2017. However, linkage analysis is not feasible for patients without affected relatives.…”

Multiple displacement amplification as the first step can increase the diagnostic efficiency of preimplantation genetic testing for monogenic disease for β‐thalassemia

“…The protocol for nested PCR was described in our previous study . The total volume of the external amplification reaction was 50 μL, including the following: 5 μL of cell lysis template fluid, 4.5 μL of 10× PCR buffer, 1.5 μL of 50 mmol/L MgCl 2 , 1 UL of 10 mmol/L deoxy‐ribonucleoside triphosphate (dNTP), 0.5 μL of β‐globin amplified primers (at a concentration of 10 mmol/L) and 0.5 U of Taq enzyme.…”

“…The procedure used for hybridization to amino‐modified oligonucleotides for β‐thalassemia, color detection and analysis of the common mutations in Hemoglobin beta locus was previously described …”

Successful clinical application of pre‑implantation genetic diagnosis for infantile neuroaxonal dystrophy

“…With the advent of next-generation sequencing (NGS) techniques, an opportunity arises to perform cost�efficient genetic testing by sequencing in different clinical scenarios, which may contribute to definitive improvements in the genetic assessment of embryos prior to transfer to the uterus (19). NGS-based methods, including Mutated Allele Revealed by Sequencing with Aneuploidy and Linkage Analyses and haplotyping analysis, have been successfully applied in SGDs and chromosomal copy number assessment (7,18,20,21).…”