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Application of single sperm sequencing for the preimplantation genetic testing of a Chinese family affected with Spinal muscular atrophy / 中华医学遗传学杂志
Chinese Journal of Medical Genetics ; (6): 148-154, 2023.
Article in Chinese | WPRIM | ID: wpr-970895
ABSTRACT
OBJECTIVE@#To assess the value of single sperm sequencing in preimplantation genetic testing for monogenic disease (PGT-M).@*METHODS@#A Chinese couple with two children whom had died of Spinal muscular atrophy (SMA) and attended the Jiangxi Provincial Maternal and Child Health Care Hospital in June 2020 was selected as the subject. Eleven single sperm samples were isolated by mechanical immobilization and subjected to whole genome amplification. Real-time PCR and Sanger sequencing were used to detect the SMN1 variants in the single sperm samples. Genomic DNA of the wife, her parents and the husband, as well as one single sperm sample harboring the SMN1 variant and two single sperm samples without the variant were used for the linkage analysis. Targeted capture and high-throughput sequencing were carried out to test 100 single nucleotide polymorphisms distributed within 2 Mb up- and downstream the variant site. The haplotypes linked with the SMN1 variants were determined by linkage analysis. Blastocyst embryos were harvested after fertilizing by intracytoplasmic sperm injection. Cells from the trophoblasts of each embryo were biopsied and subjected to whole genome amplification and targeted capture and high-throughput sequencing to determine their carrier status. Chromosomal aneuploidy of wild-type embryos was excluded. An euploid embryo of high quality was transferred. Amniotic fluid sample was taken at 18 weeks of gestation to confirm the status of the fetus.@*RESULTS@#Genetic testing showed that the couple both had deletion of exons 7 ~ 8 of the SMN1 gene. The wife has inherited the deletion from her father, while the husband was de novo. The haplotypes of the husband were successfully constructed by single sperm sequencing. Preimplantation genetic testing has indicated that 5 embryos had harbored the heterozygous variant, 4 embryos were of the wild type, among which 3 were euploid. Prenatal diagnosis during the second trimester of pregnancy has confirmed that the fetus did not carry the deletion.@*CONCLUSION@#By single sperm sequencing and PGT-M, the birth of further affected child has been successfully avoided.
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Semen / Spermatozoa / Blastocyst / Muscular Atrophy, Spinal / Genetic Testing / Preimplantation Diagnosis / High-Throughput Nucleotide Sequencing / East Asian People / Aneuploidy Limits: Child / Female / Humans / Male / Pregnancy Language: Chinese Journal: Chinese Journal of Medical Genetics Year: 2023 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Semen / Spermatozoa / Blastocyst / Muscular Atrophy, Spinal / Genetic Testing / Preimplantation Diagnosis / High-Throughput Nucleotide Sequencing / East Asian People / Aneuploidy Limits: Child / Female / Humans / Male / Pregnancy Language: Chinese Journal: Chinese Journal of Medical Genetics Year: 2023 Type: Article