Parks Michael, Court Samantha, Cleary Siobhan, Clokie Samuel, Hewitt Julie, Williams Denise, Cole Trevor, MacDonald Fiona, Griffiths Mike, Allen Stephanie
West Midlands Regional Genetics Laboratory, Birmingham Women's NHS Foundation Trust, Birmingham, UK.
Prenat Diagn. 2016 Apr;36(4):312-20. doi: 10.1002/pd.4781. Epub 2016 Feb 23.
Development of an accurate and affordable test for the non-invasive prenatal diagnosis of Duchenne and Becker muscular dystrophies (DMD/BMD) to implement in clinical practice.
Cell-free DNA was extracted from maternal blood and prepared for massively parallel sequencing on an Illumina MiSeq by targeted capture enrichment of single nucleotide polymorphisms (SNPs) across the dystrophin gene on chromosome X. Sequencing data were analysed by relative haplotype dosage.
Seven healthy pregnant donors and two pregnant DMD carriers all bearing a male fetus were recruited through the non-invasive prenatal diagnosis for single gene disorders study. Non-invasive prenatal diagnosis testing was conducted by relative haplotype dosage analysis for X-linked disorders where the genomic DNA from the chorionic villus sampling (for healthy pregnant donors) or from the proband (for pregnant DMD carriers) was used to identify the reference haplotype. Results for all patients showed a test accuracy of 100%, when the calculated fetal fraction was >4% and correlated with known outcomes. A recombination event was also detected in a DMD patient.
Our new test for NIPD of DMD/BMD has been shown to be accurate and reliable during initial stages of validation. It is also feasible for implementation into clinical service.
开发一种准确且经济实惠的检测方法，用于杜氏和贝克型肌营养不良症（DMD/BMD）的无创产前诊断，以便在临床实践中应用。
从母血中提取游离DNA，并通过对X染色体上肌营养不良蛋白基因的单核苷酸多态性（SNP）进行靶向捕获富集，为在Illumina MiSeq上进行大规模平行测序做准备。测序数据通过相对单倍型剂量进行分析。
通过单基因疾病的无创产前诊断研究招募了7名健康的怀孕捐赠者和2名怀有男性胎儿的怀孕DMD携带者。对于X连锁疾病，通过相对单倍型剂量分析进行无创产前诊断检测，其中来自绒毛取样（针对健康怀孕捐赠者）或先证者（针对怀孕DMD携带者）的基因组DNA用于确定参考单倍型。当计算出的胎儿游离DNA比例>4%且与已知结果相关时，所有患者的检测准确率均为100%。在一名DMD患者中还检测到了一次重组事件。
我们新的DMD/BMD无创产前诊断检测方法在验证的初始阶段已被证明准确可靠。将其应用于临床服务也是可行的。
