X連鎖先天性視網(wǎng)膜劈裂XLRS1基因突變檢測(cè)
發(fā)布時(shí)間：2018-10-05 08:16
【摘要】：目的： 對(duì)三個(gè)視網(wǎng)膜劈裂家系(包括4例患者)從基因水平檢測(cè)XLRS1基因突變位點(diǎn)及類(lèi)型,并為患者及家屬提供遺傳咨詢(xún)。 方法： 對(duì)視網(wǎng)膜劈裂患者進(jìn)行全面的眼科檢查包括最佳矯正視力、裂隙燈顯微鏡檢查、散瞳后眼底檢查及彩色眼底照相、光學(xué)相關(guān)斷層掃描(optical coherence tomography, OCT),視網(wǎng)膜電圖(electroretinogram, ERG)檢查和熒光素血管造影(fundus angiography,FFA)。對(duì)家系內(nèi)成員進(jìn)行最佳矯正視力、裂隙燈顯微鏡檢查、眼底檢查,必要時(shí)行OCT,FFA,ERG等檢查。采集患者及家系成員外周血,提取DNA,采用聚合酶鏈反應(yīng)-單鏈構(gòu)象多態(tài)性(polymerase chain reaction-single strand conformation polymorphism, PCR-SSCP)方法,對(duì)XLRS1基因的6個(gè)外顯子各片段進(jìn)行擴(kuò)增,擴(kuò)增區(qū)域囊括XLRS1基因的全部編碼區(qū),應(yīng)用單鏈構(gòu)像多態(tài)性(single strand conformation polymorphism, SSCP)分析,對(duì)PCR產(chǎn)物進(jìn)行DNA測(cè)序,將測(cè)序結(jié)果與XLRS1標(biāo)準(zhǔn)序列比對(duì)(MIM：312700),以明確突變位點(diǎn)及突變類(lèi)型。 結(jié)果： 確診為視網(wǎng)膜劈裂的4名患者來(lái)源于三個(gè)家系,均為男性,符合X連鎖遺傳特征,患者年齡為8歲～16歲之間,平均10.75歲,平均視力0.22,OCT表現(xiàn)為視網(wǎng)膜囊樣改變。ERG表現(xiàn)為b波下降,a波輕度下降或正常。經(jīng)DNA測(cè)序后發(fā)現(xiàn)2種不同類(lèi)型的致病基因突變,第一種為堿基替換導(dǎo)致錯(cuò)義突變：家系1基因突變?yōu)閏.625CT(p.R209C),家系3基因突變?yōu)閏.002TC(p.M1T),導(dǎo)致合成的蛋白質(zhì)功能異常；第二種為堿基缺失：家系2患者第1外顯子未能擴(kuò)增出,懷疑第一外顯子缺失,第2-6外顯子檢測(cè)無(wú)異常；在家系1家庭成員中發(fā)現(xiàn)3名女性攜帶者,家系3家庭成員中發(fā)現(xiàn)5名女性攜帶者。 結(jié)論： 1.本研究從基因水平明確了XLRS(?)的診斷,并發(fā)現(xiàn)這些患者的XLRS1基因突變或缺失。 2.本研究確診了8名XLRS1基因攜帶者,對(duì)各個(gè)家系的遺傳咨詢(xún)及生育指導(dǎo)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.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類(lèi)號(hào)】：R774.1
本文編號(hào)：2252660
[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.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類(lèi)號(hào)】：R774.1
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，本文編號(hào)：2252660
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