中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2011年
5期
427-431
,共5页
鞠宏%李宁东%赵堪兴%王犁明%王玉川%应铭%高翔
鞠宏%李寧東%趙堪興%王犛明%王玉川%應銘%高翔
국굉%리저동%조감흥%왕리명%왕옥천%응명%고상
先天性白内障%基因连锁分析%遗传
先天性白內障%基因連鎖分析%遺傳
선천성백내장%기인련쇄분석%유전
congenital cataract%Linkage analysis%fluorescent labeling%universal M13
背景 先天性白内障约1/3的病例是由遗传所致,已发现遗传性白内障有着极为明显的遗传异质性,了解先天性白内障的致病基因对其基因治疗极为重要.目的 分析一个具有常染色体显性遗传特点的先天性白内障家系的临床表型特征.进行已知致病基因的筛查定位.方法对遗传性先天性白内障一家系共16名成员眼部进行详细的临床检查,包括6例患者,确定为本家系白内障患者的临床表型.收集其中11名家系成员的血液样本提取DNA,包括3名正常家系成员及其配偶、5例患者.利用连锁分析进行排除定位,并采用Schuelke报道的新方法,只合成普通引物及一种荧光标记的通用引物M13,进行聚合酶链反应(PCR),对连锁区域内的候选基因进行基因序列分析.结果 本家系的白内障遗传方式符合常染色体显性遗传特征.基因连锁分析表明,在D22S315得到最高LOD值为1.20,在D16S3068得到LOD值为0.6.CRYBB2基因所有编码区及外显子与内含子交界处未发现基因序列突变.结论 本家系初步排除了CRYBB2基因与此家系先天性白内障的相关性.对这个家系的基因定位需要更进一步的全基因组扫描,以发现致病基因在染色体上的可疑区间.连锁分析中进行微卫星位点的PCR扩增时,利用合成荧光标记的通用引物M13.可以显著降低成本,并取得同样的实验结果.
揹景 先天性白內障約1/3的病例是由遺傳所緻,已髮現遺傳性白內障有著極為明顯的遺傳異質性,瞭解先天性白內障的緻病基因對其基因治療極為重要.目的 分析一箇具有常染色體顯性遺傳特點的先天性白內障傢繫的臨床錶型特徵.進行已知緻病基因的篩查定位.方法對遺傳性先天性白內障一傢繫共16名成員眼部進行詳細的臨床檢查,包括6例患者,確定為本傢繫白內障患者的臨床錶型.收集其中11名傢繫成員的血液樣本提取DNA,包括3名正常傢繫成員及其配偶、5例患者.利用連鎖分析進行排除定位,併採用Schuelke報道的新方法,隻閤成普通引物及一種熒光標記的通用引物M13,進行聚閤酶鏈反應(PCR),對連鎖區域內的候選基因進行基因序列分析.結果 本傢繫的白內障遺傳方式符閤常染色體顯性遺傳特徵.基因連鎖分析錶明,在D22S315得到最高LOD值為1.20,在D16S3068得到LOD值為0.6.CRYBB2基因所有編碼區及外顯子與內含子交界處未髮現基因序列突變.結論 本傢繫初步排除瞭CRYBB2基因與此傢繫先天性白內障的相關性.對這箇傢繫的基因定位需要更進一步的全基因組掃描,以髮現緻病基因在染色體上的可疑區間.連鎖分析中進行微衛星位點的PCR擴增時,利用閤成熒光標記的通用引物M13.可以顯著降低成本,併取得同樣的實驗結果.
배경 선천성백내장약1/3적병례시유유전소치,이발현유전성백내장유착겁위명현적유전이질성,료해선천성백내장적치병기인대기기인치료겁위중요.목적 분석일개구유상염색체현성유전특점적선천성백내장가계적림상표형특정.진행이지치병기인적사사정위.방법대유전성선천성백내장일가계공16명성원안부진행상세적림상검사,포괄6례환자,학정위본가계백내장환자적림상표형.수집기중11명가계성원적혈액양본제취DNA,포괄3명정상가계성원급기배우、5례환자.이용련쇄분석진행배제정위,병채용Schuelke보도적신방법,지합성보통인물급일충형광표기적통용인물M13,진행취합매련반응(PCR),대련쇄구역내적후선기인진행기인서렬분석.결과 본가계적백내장유전방식부합상염색체현성유전특정.기인련쇄분석표명,재D22S315득도최고LOD치위1.20,재D16S3068득도LOD치위0.6.CRYBB2기인소유편마구급외현자여내함자교계처미발현기인서렬돌변.결론 본가계초보배제료CRYBB2기인여차가계선천성백내장적상관성.대저개가계적기인정위수요경진일보적전기인조소묘,이발현치병기인재염색체상적가의구간.련쇄분석중진행미위성위점적PCR확증시,이용합성형광표기적통용인물M13.가이현저강저성본,병취득동양적실험결과.
Background About one third of congenital cataract is associated with inheriting factor.The inherited heterogeneity has been found in congenital cataract.To seek the pathogenic gene is essential for the gene therapy. Objective Present study was to map and identify the causal gene for autosomal dominant congenital cataract (ADCC) in a Chinese family. Methods The clinical features of all affected members in this family were examined.Blood samples were collected from eleven family members for genetic linkage analysis.Polymorphic microsatellite markers were selected from the regions which harbor all known loci linked with ADCC.Universal fluorescent-labeled M13 primer was used in linkage analysis.Direct genomic sequencing was used to evaluate the candidate gene for example CRYBB2 gene.This study followed Helsinki Declaration and was proved by Tianjin City Ethic Committee.Written informed consent was obtained from each SUbject before any medical procees. Results The maximum two-point LOD score of 1.20 was obtained for marker D22S315 (θ=0).The LOD score of 0.6 was obtained for marker D16S3068.No mutation in all exons of CRYBB2 gene was found in the family. Conclusion CRYBB2 gene associated with ADCC was excluded from the family.A genome-wide linkage screening should be conducted.Genotyping with microsatellite markers using Universal fluorescent-labeled M13 primer can decrease the cost and obtain the same result.