【摘要】 目的 探讨基于二代测序（NGS）技术的单核苷酸多态性（SNP）连锁分析在常染色体隐性遗传性多囊肾病（ARPKD）家系胚胎植入前遗传学检测（PGT）中的应用价值。方法 选取1个ARPKD家系，女方孕期产检发现胎儿有多囊肾行引产，胎儿基因检测为多囊肾/多囊肝病变1基因（PKHD1）复合杂合突变c.10444C> T（父源）和c.4303del（母源），其中c.4303del突变为首次报道。采用多重聚合酶链反应（PCR）和NGS在突变位点两侧2M区域内选择335个信息丰富、紧密连锁的SNP位点作为遗传连锁标记，构建夫妇双方携带基因突变的SNP风险单体型。体外受精后行囊胚培养。对活检获得的滋养层细胞进行全基因组扩增（WGA）后，采用Sanger测序直接检测PKHD1基因突变，采用基于NGS的SNP连锁分析鉴别携带突变的染色体，同时进行拷贝数变异（CNV）分析以进行低深度的染色体非整倍性筛查。结果 6个活检囊胚中有4个为未携带突变的整倍体，有1个携带杂合突变的嵌合体，1个测序数据波动大，无法判断。选择其中1枚未检测到突变的优质整倍体胚胎进行冻融胚胎移植（FET），足月分娩一健康婴儿。结论 应用基于NGS的SNP连锁分析进行PGT具有高效稳定的特点，可有效阻断ARPKD在家系中的垂直传递，同时可避免因妊娠非整倍体胚胎而导致的流产问题。该研究也是首次针对PKHD1基因c.4303del突变的PGT报道。更多还原

【Abstract】 Objective To investigate the application value of single nucleotide polymorphism(SNP) linkage analysis based on next-generation sequencing(NGS) technology in preimplantation genetic testing(PGT) of families with autosomal recessive polycystic kidney disease(ARPKD). Methods A family with ARPKD was selected, where the female member had a pregnancy ultrasound revealing polycystic kidney in the fetus. Genetic testing showed compound heterozygous mutations of the polycystic kidney/polycystic liver disease 1 gene(PKHD1), c.10444C > T(paternal) and c.4303del(maternal), with the c.4303del mutation being reported for the first time.Targeting the coding region of the PKHD1 gene, 335 high-density tightly linked SNP sites were selected in the upstream and downstream 2M regions using multiplex polymerase chain reaction(PCR) and NGS. The couple’s SNP risk haplotypes carrying gene mutations were constructed. After in vitro fertilization, blastocyst culture was performed. Trophoblastic cells obtained from the biopsy were subjected to whole-genome amplification, and NGS was used for linkage analysis and low-depth chromosomal aneuploidy screening of the embryos. Sanger sequencing was used to verify the results of embryo linkage analysis. Results Among the 6 biopsied embryos, 4 were mutation-free and euploid, 1 exhibited heterozygous for the mutation and mosaic while another unstable sequencing data, making it impossible to judge. One of the mutation-free and developmentally healthy euploid embryos was implanted into the maternal uterus, resulting in the full-term delivery of a healthy baby. Conclusion Application of NGS-based SNP linkage analysis in PGT can effectively blocking the vertical transmission of ARPKD within families, while avoiding abortion issues caused by aneuploid embryos. This study is also the first PGT report targeting the PKHD1 gene c.4303del mutation.更多还原