论文部分内容阅读
目的对中国人先天性眼外肌纤维化(CFEOM)家系和散发个体进行致病基因突变筛查。设计实验研究。研究对象6个CFEOM家系和4个CFEOM散发个体。方法收集CFEOM患者的临床及眼运动神经MRI检查资料,采集患者及家庭成员外周静脉血,提取基因组DNA,应用目标序列捕获联合高通量测序的方法对11个疾病候选基因进行全部外显子的突变筛查。生物信息学分析得到候选基因碱基改变后,在100个无关正常对照人群中进行PCR和Sanger测序验证并对基因型和表型特征进行分析。主要指标基因序列。结果 80%(8/10)CFEOM先证者存在KIF21A基因突变,分别为c.2860C>T(p.R954W)和c.2861G>A(p.R954Q);20%(2/10)先证者存在TUBB3基因突变,分别为c.784C>T(p.R262C)和c.1138C>T(p.R380C)。所有突变在家系正常个体及正常对照人群中均未检测到。携带KIF21A基因突变的患者表现为CFEOM1A和CFEOM3B表型,而携带TUBB3基因突变的患者表现出CFEOM3A表型。结论 KIF21A基因突变是中国人CFEOM的常见致病原因。高通量基因筛查可快速、准确地协助疾病的临床诊断和基因分型。
Objective To screen for mutations of causative genes in Chinese pedigrees and individuals with congenital extraocular muscle fibrosis (CFEOM). Design experiment research. Subjects Six CFEOM pedigrees and four CFEOM individuals were distributed. Methods The clinical and oculomotor nerve MRI examination data of CFEOM patients were collected. Peripheral venous blood of patients and their family members were collected and genomic DNA was extracted. The target sequences were captured and combined with high-throughput sequencing to detect 11 disease candidate genes in all exon Mutation screening. After bioinformatics analysis, the base of candidate gene was changed, PCR and Sanger sequencing were performed in 100 unrelated normal controls and the genotype and phenotypic characteristics were analyzed. The main indicator gene sequence. Results 80% (8/10) CFEOM probands had KIF21A gene mutations, which were c.2860C> T (p.R954W) and c.2861G> A (p.R954Q) respectively; 20% (2/10) There were TUBB3 gene mutations, namely c.784C> T (p.R262C) and c.1138C> T (p.R380C). All mutations were not detected in normal pedigrees and normal control subjects. Patients carrying mutations in the KIF21A gene exhibited the CFEOM1A and CFEOM3B phenotypes while those carrying the TUBB3 gene mutation showed the CFEOM3A phenotype. Conclusion KIF21A gene mutation is a common cause of CFEOM in Chinese. High-throughput gene screening can quickly and accurately assist in the clinical diagnosis and genotyping of diseases.