国际生殖健康/计划生育杂志2017，Vol.36Issue(4)：276-279,封3,5.
X-连锁慢性肉芽肿病的植入前遗传学诊断方法学研究
Methodology Study on Preimplantation Genetic Diagnosis of X-linked Chronic Granulomatous Disease
摘要
Abstract
Objective:To develop a reliable and exact protocol for the multiple displacement amplification (MDA)-based preimplantation genetic diagnosis (PGD) of X-linked chronic granulomatous disease (X-CGD).Methods:The haplotype analysis of X-CGD was performed for 16 short tandem repeats (STR) loci on both sides of the CYBB gene.Whole-genome amplification of single cell was carried out by MDA.The STR loci with polymorphism as demonstrated by the linkage analysis and the specific amplification of CYBB gene were analyzed.The locus Amel was used for sex determination.Results:Seven STR loci were found to be polymorphic in the linkage analysis.MDA was performed in 10 single lymphocytes and 10 single blastomeres.The efficiencies of PCR amplification of the pathogenic CYBB exon 8 was 100％.ADO of the mutant allele of CYBB was detected in one of the lymphocytes,the ADO rate 10％ (1/10).For those single blastomeres,the 215 bp band corresponding to the normal allele was detected in all MDA products,while the abnormal 204 bp band was not detected.For the 7 polymorphic STR loci and AMELs,the PCR amplification efficiency and the ADO rote were 96.9％ (155/160) and 11.3％ (13/115),respectively.Conclusions:In this study,MDA combined with pathogenic gene specific amplification and haplotype analysis was performed to detect chronic granulomatous disease in a single cell level.The combination of the two techniques can improve the efficiency of PGD for CGD by reducing the misdiagnosis caused by contamination and ADO.关键词
X-连锁慢性肉芽肿病/系谱/植入前诊断/多重置换扩增/单体型分析Key words
X-linked chronic granulomatous disease/Pedigree/Preimplantation diagnosis/Multiple displacement amplification/Haplotype analysis引用本文复制引用
吴海涛,沈晓婷,刘瑜亮,钟依平,王静,曾艳红,丁晨晖,周灿权..X-连锁慢性肉芽肿病的植入前遗传学诊断方法学研究[J].国际生殖健康/计划生育杂志,2017,36(4):276-279,封3,5.基金项目
国家自然科学基金(81370765),广州市科技计划项目(201300000097),广东省科技计划项目(2013B021800271) （81370765）
