一个F8基因大片段缺失的血友病A家系遗传学分析  

Genetic analysis for a family affected with hemophilia type A due to a large deletion of F8 gene

作　　者：白楠[1] 梅世月[1] 刘宁[1] 赵振华[1] 孟静静[1] 孔祥东[1] 

机构地区：[1]郑州大学第一附属医院产前诊断中心,450052

出　　处：《中华医学遗传学杂志》2016年第6期782-785,共4页Chinese Journal of Medical Genetics

摘　　要：目的检测1个重型血友病A（hemophilia A，HA）患者家系凝血因子Ⅷ基因（F8）的突变情况，明确F8基因的突变类型，为家系提供遗传咨询。方法联合应用反向转变PCR（inverse-shifting PCR,，IS-PCR）、二代测序技术（next generation sequencing, NGS）、多重连接探针扩增技术（ multiplex ligation dependent probe amplification, MLPA）和短串联重复序列（short tandem repeat,STR）连锁分析等方法对1个重型血友病A家系先证者、携带者及胎儿进行分析。结果IS-PCR结果显示先证者未发生F8基因第1内含子倒位（intron 1 inversion, Inv1）或第22内含子倒位（intron 22 inversion，Inv22）；NGS结果显示先证者船基因未发生点突变以及小的插入缺失，但在NGS覆盖度分析中发现先证者F8基因第2外显子测序深度为0；MLPA结果显示先证者F8-E2缺失，先证者母亲为F8基因第2外显子杂合缺失携带者。另外连锁分析与其它几种方法相比较，出现了结果不一致的情况。结论通过NGS确定1例F8基因第2外显子大片段缺失突变，血友病A家系F8基因连锁分析用于携带者筛查及产前诊断需慎重，多种方法联合分析可准确判断F8基因的突变情况。Objective To detect potential mutation of F8 gene in a family affected with hemophilia type A. Methods Inverse-shifting PCR （IS-PCR）, next-generation sequencing （NGS）, multiplex ligation- dependent probe amplification （MLPA）, and short tandem repeat （STR） assays were used. Results IS- PCR showed that no inversion of F8 gene has occurred in the family. NGS detected no point mutation or small InDel in the proband, but suggested that the exon 2 of the F8 gene may be deleted. MLPA also showed that exon 2 of the F8 gene was absent in the proband, while the carriers were heterozygous for the deletion, though STR analysis yielded a paradoxical result. Conclusion NGS analysis has identified a large deletion of exon 2 of the F8 gene in a family affected with hemophilia A. Discretion is required when STR analysis was used for carrier screening and antenatal diagnosis. Combination of multiple methods can improve the accuracy for the detection of F8 gene mutations.

关 键 词：反向转变PCR 二代测序技术 多重连接探针扩增技术 外显子缺失 

分 类 号：R5[医药卫生—内科学]