【摘要】 目的利用目标基因捕获高通量测序技术和Sanger测序方法,对三个先天性晶状体半脱位家系患者FBN1和ADAMTSL4基因进行突变筛查。方法提取三个家系14例晶状体半脱位患者及3名家系正常成员外周静脉血全基因组DNA,利用目标基因捕获技术,设计FBN1的65个外显子区域捕获探针,与基因组DNA文库混合杂交,收集洗脱处理后的目标基因组DNA,再利用Hi Seq2000进行高通量测序,通过数据分析,确定突变位点,用Sanger法验证测序结果;同时利用Sanger法对3名先证者ADAMTSL4基因进行测序分析。结果通过高通量测序和Sanger法验证,分别发现3个家系3个不同的FBN1基因编码区的杂合错义突变,通过Sanger测序法发现4个ADAMTSL4基因编码区的单核苷酸多态性改变。结论 3个先天性晶状体半脱位家系的致病性基因为FBN1基因,FBN1基因的突变是导致患者晶状体半脱位的临床表型的主要原因。更多还原

【Abstract】 Objective To screen the genetic mutations in FBN1 and ADAMTSL4 genes from three families who had congenital lens subluxation by targeted gene capture w ith high-throughput and Sanger sequencing techniques.Methods Peripheral blood samples w ere collected from 14 lens subluxation patients and 3 healthy members of three families and genomic DNA w as isolated. FBN1 w as selected by a gene capture strategy w ith 65 exon capture probes. The enrichment libraries w ere sequenced on Hi Seq2000 sequencer to determine the mutation frequency in FBN1. Probable mutations w ere verified by Sanger sequencing method. At the same time,ADAMTSL4 gene from three probands w as also analyzed by Sanger method. Results Three different hybrid missense mutations w ere found in FBN1 gene from the three families. Four single nucleotide polymorphism changes w ere found in ADAMTSL4 gene. Conclusion FBN1 gene is the pathogenic gene that cause congenital lens subluxation in the selected three families. FBN1 genetic mutations are the primary causes leading to the clinical signs of lens subluxation.更多还原