基于单体型的杜氏肌营养不良无创产前检测研究

Haplotype-Based Noninvasive Prenatal Testing for Duchenne Muscular Dystrophy

目的:探讨利用先证者辅助单体型分析方法(PAHP)对杜氏肌营养不良(DMD)进行无创产前检测(NIPT)的可行性。方法:招募生育过DMD先证者并再次妊娠的家系17例。对孕妇、孕妇丈夫与先证者外周血基因组DNA(g DNA)样本进行目标序列捕获测序,获得孕妇与致病位点连锁的单体型信息。然后在夫妻双方单体型的辅助下,通过对血浆数据中各个信息可供单核苷酸多态性(SNP)位点分析及统计,推断在DMD基因区域胎儿获得的母源单体型是否与先证者一致。携带与先证者相同单体型的男胎为患胎,女胎为携带者,其余为正常胎。将NIPT结果与DMD基因诊断金标准进行比较,验证其准确性。结果:NIPT结果提示17个家系中9例为男性胎儿,8例为女性胎儿;患胎3例,携带者4例,正常胎10例。该结果与胎儿的DMD基因诊断结果一致,无假阳性与假阴性结果。结论:基于先证者辅助单体型分析方法的NIPT取材方便,能避免宫内介入性操作,同时结果准确,应用于DMD的产前检测具有一定的可行性。

Objective： To explore the feasibility of a haplotype-based noninvasive prenatal testing（ NIPT） for Duchenne Muscular Dystrophy（ DMD）.Methods： Seventeen families,each with a proband affected by DMD were recruited.Capture sequencing was performed on genomic DNA in peripheral blood from parents and proband.Maternal plasma DNA also underwent targeted sequencing to deduce fetal haplotypes assisted,which by parental haplotypes using a hidden Markov model.The fetal genotypes in DMD gene were further validated by invasive procedures of prenatal diagnosis. Results： The haplotype-based NIPT was successfully performed in all families. Nine male and eight female fetuses were identified.Four fetuses were carriers and three male fetuses were diagnosed with DMD patients. All these results were consistent with those of invasive procedures. Conclusions： Haplotypebased noninvasive prenatal testing for DMD targeted sequencing is safe and accurate and has potential for clinical implementation.