X连锁先天性视网膜劈裂XLRS1基因突变检测

发布时间：2018-10-05 08:16

【摘要】：目的：对三个视网膜劈裂家系(包括4例患者)从基因水平检测XLRS1基因突变位点及类型,并为患者及家属提供遗传咨询。方法：对视网膜劈裂患者进行全面的眼科检查包括最佳矫正视力、裂隙灯显微镜检查、散瞳后眼底检查及彩色眼底照相、光学相关断层扫描(optical coherence tomography, OCT),视网膜电图(electroretinogram, ERG)检查和荧光素血管造影(fundus angiography,FFA)。对家系内成员进行最佳矫正视力、裂隙灯显微镜检查、眼底检查,必要时行OCT,FFA,ERG等检查。采集患者及家系成员外周血,提取DNA,采用聚合酶链反应-单链构象多态性(polymerase chain reaction-single strand conformation polymorphism, PCR-SSCP)方法,对XLRS1基因的6个外显子各片段进行扩增,扩增区域囊括XLRS1基因的全部编码区,应用单链构像多态性(single strand conformation polymorphism, SSCP)分析,对PCR产物进行DNA测序,将测序结果与XLRS1标准序列比对(MIM：312700),以明确突变位点及突变类型。结果：确诊为视网膜劈裂的4名患者来源于三个家系,均为男性,符合X连锁遗传特征,患者年龄为8岁～16岁之间,平均10.75岁,平均视力0.22,OCT表现为视网膜囊样改变。ERG表现为b波下降,a波轻度下降或正常。经DNA测序后发现2种不同类型的致病基因突变,第一种为碱基替换导致错义突变：家系1基因突变为c.625CT(p.R209C),家系3基因突变为c.002TC(p.M1T),导致合成的蛋白质功能异常；第二种为碱基缺失：家系2患者第1外显子未能扩增出,怀疑第一外显子缺失,第2-6外显子检测无异常；在家系1家庭成员中发现3名女性携带者,家系3家庭成员中发现5名女性携带者。结论： 1.本研究从基因水平明确了XLRS(?)的诊断,并发现这些患者的XLRS1基因突变或缺失。 2.本研究确诊了8名XLRS1基因携带者,对各个家系的遗传咨询及生育指导有重要作用。
[Abstract]:Objective: to detect the mutation site and type of XLRS1 gene at gene level in three families (including 4 patients), and to provide genetic counseling for patients and their families. Methods: a comprehensive ophthalmic examination including best corrected visual acuity, slit lamp microscope, fundus examination after pupil dilation and color fundus photography were performed in patients with retinal splitting. Optical correlation tomography (optical coherence tomography, OCT),) electroretinogram (electroretinogram, ERG) and fluorescein angiography (fundus angiography,FFA). Best corrected visual acuity, slit lamp microscope, fundus examination, OCT,FFA,ERG, etc. DNA, was extracted from peripheral blood of patients and family members and amplified by polymerase chain reaction-single strand conformation polymorphic (polymerase chain reaction-single strand conformation polymorphism, PCR-SSCP (polymerase chain reaction-single strand conformation polymorphism, PCR-SSCP) method. The six exons of the XLRS1 gene were amplified and the regions were amplified to include all the coding regions of the XLRS1 gene. Single strand conformation polymorphism (SSCP) (single strand conformation polymorphism, SSCP) analysis was used to sequence the PCR products. The results were compared with the XLRS1 standard sequence (MIM:312700) to identify the mutation sites and mutation types. Results: the 4 patients who were diagnosed as retinoschisis were from three families, all of them were male. The patients were between 8 and 16 years old, with an average age of 10.75 years. The mean visual acuity of 0.22 OCT showed retinal cystoid changes. ERG showed a slight decrease of b wave or a wave. After DNA sequencing, two different types of pathogenetic gene mutations were found. The first was base substitution, which led to missense mutation: family 1 gene mutation to c.625CT (p.R209C), pedigree 3 gene mutation to c.002TC (p.M1T), which led to abnormal function of synthesized protein. The second was base deletion: the first exon was not amplified in the family 2 patients, the deletion of the first exon was suspected and the exon 2-6 was not abnormal. 3 female carriers were found in the family members of family 1. Five female carriers were found in family members of family 3. Conclusion: 1. In this study, XLRS (?) XLRS1 gene mutations or deletions were found in these patients. 2. This study confirmed 8 carriers of XLRS1 gene and played an important role in genetic counseling and fertility guidance.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R774.1

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