Application of polar body sequencing for preimplantation genetic testing of a female patient with Van der Woude syndrome
10.3760/cma.j.cn101441-20211014-00460
- VernacularTitle:极体测序技术在Van der Woude综合征患者胚胎植入前遗传学检测中的应用
- Author:
Jia CHEN
1
;
Xingwu WU
;
Lifeng TIAN
;
Ge CHEN
;
Zhihui HUANG
;
Cailin XIN
;
Yan ZHAO
;
Yanqiu LIU
;
Qiongfang WU
Author Information
1. 江西省妇幼保健院生殖医学中心，南昌 330006
- Publication Type:Journal Article
- Keywords:
Van der Woude syndrome;
De novo variants;
Preimplantation genetic testing;
Polar body sequencing;
Allele drop out
- From:
Chinese Journal of Reproduction and Contraception
2023;43(1):78-83
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the value of polar body sequencing in preimplantation genetic testing (PGT) for monogenic disease of a female patient with Van der Woude syndrome.Methods:PGT based on polar body sequencing was performed for a female patient with Van der Woude syndrome caused by a de novoIRF6 pathogenic variant. Totally six oocytes were fertilized by intracytoplasmic sperm injection (ICSI). The first, second polar bodies and the trophoblast ectoderm cells of blastocysts were biopsied respectively. Sanger sequencing was used to detect the pathogenic variant in the biopsied cells after genome-wide amplification. The genotypes and pathogenic possibilities of the embryos were inferred according to the genotypes of corresponding tested polar bodies. In order to prevent the absence of transplantable embryos due to the failure of blastocyst culture, vitrification was performed on an embryo with good morphology and low pathogenic possibility before blastocyst formation. The 175 single nucleotide polymorphisms (SNPs) within the 1M region upstream and downstream from the pathogenic variant location were tested by targeted capture sequencing in the couple and selected polar bodies and embryos to construct the haplotypes. An embryo with low pathogenic possibility was transferred. Prenatal diagnosis was strongly recommended after successful pregnancy. Prenatal and postnatal follow-up were performed. Results:Totally six first polar bodies and six second polar bodies were obtained. The pathogenic variant was successfully sequenced in 11 polar bodies. Among the six embryos, one embryo with low pathogenic possibility was vitrified on day 4 (D4) after fully informed consent of the couple; one embryo developed to blastocyst was detected with high pathogenic possibility; the other four embryos were degenerated during blastocyst culture. The SNP haplotypes closely linked to the pathogenic variant location were successfully constructed by linkage analysis. The haplotype analysis of the embryos was in consistent with Sanger sequencing. The D4 embryo with low pathogenic possibility was transferred. The couple refused to conduct invasive prenatal diagnosis after pregnancy. None orofacial clefts were detected after the baby was born, and the pathogenic variant was not detected in the neonatal cord blood either.Conclusion:This study successfully blocked a female patient with Van der Woude syndrome caused by a de novoIRF6 pathogenic variant give birth to an affected baby by polar body sequencing based preimplantation genetic testing for monogenic disease.
