通过单细胞测序对β地中海贫血家族进行胚胎植入前基因诊断的胚胎基因组分析。
Embryo genome profiling by single-cell sequencing for preimplantation genetic diagnosis in a β-thalassemia family.
作者信息
Xu Yanwen, Chen Shengpei, Yin Xuyang, Shen Xiaoting, Pan Xiaoyu, Chen Fang, Jiang Hui, Liang Yu, Wang Wei, Xu Xun, Wang Jian, Zhang Xiuqing, Zhou Canquan, Wang Jun
机构信息
First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China; Guangdong Provincial Key Laboratory of Reproductive Medicine, Guangdong, China;
BGI-Shenzhen, Shenzhen, China; Shenzhen Municipal Key Laboratory of Birth Defects Screening and Engineering, Shenzhen, China; Guangdong Provincial Key Laboratory of Human Diseases Genome, Guangdong, China; State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, China;
出版信息
Clin Chem. 2015 Apr;61(4):617-26. doi: 10.1373/clinchem.2014.228569. Epub 2015 Feb 26.
BACKGROUND
The embryonic genome, including genotypes and haplotypes, contains all the information for preimplantation genetic diagnosis, representing great potential for mendelian disorder carriers to conceive healthy babies.
METHODS
We developed a strategy to obtain the full embryonic genome for a β-thalassemia-carrier couple to have a healthy second baby. We carried out sequencing for single blastomere cells and the family trio and further developed the analysis pipeline, including recovery of the missing alleles, removal of the majority of errors, and phasing of the embryonic genome.
RESULTS
The final accuracy for homozygous and heterozygous single-nucleotide polymorphisms reached 99.62% and 98.39%, respectively. The aneuploidies of embryos were detected as well. Based on the comprehensive embryonic genome, we effectively performed whole-genome mendelian disorder diagnosis and human leukocyte antigen matching tests.
CONCLUSIONS
This retrospective study in a β-thalassemia family demonstrates a method for embryo genome recovery through single-cell sequencing, which permits detection of genetic variations in preimplantation genetic diagnosis. It shows the potential of single-cell sequencing technology in preimplantation genetic diagnosis clinical practices.
背景
胚胎基因组，包括基因型和单倍型，包含了植入前基因诊断所需的所有信息，这为孟德尔疾病携带者孕育健康婴儿提供了巨大潜力。
方法
我们开发了一种策略，用于为一对β地中海贫血携带者夫妇获取完整的胚胎基因组，以生育一个健康的二胎宝宝。我们对单个卵裂球细胞和三联体家庭进行了测序，并进一步开发了分析流程，包括缺失等位基因的恢复、大部分错误的去除以及胚胎基因组的定相。
结果
纯合和杂合单核苷酸多态性的最终准确率分别达到99.62%和98.39%。同时也检测到了胚胎的非整倍体情况。基于完整的胚胎基因组，我们有效地进行了全基因组孟德尔疾病诊断和人类白细胞抗原匹配测试。
结论
这项在β地中海贫血家族中的回顾性研究展示了一种通过单细胞测序恢复胚胎基因组的方法，该方法能够在植入前基因诊断中检测基因变异。它显示了单细胞测序技术在植入前基因诊断临床实践中的潜力。
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