双特异探针技术早期检测先天性白内障晶状体蛋白突变基因的研究  

Early detection using dual-specific probe technique for genetic mutation of congenital cataract lens protein

作　　者：曹雨娟 邵珺[2] 虞瑛青 姚勇[2] 朱靖[2] 田玉景 曹葭 Yu-Juan Cao;Jun Shao;Ying-Qing Yu;Yong Yao;Jing Zhu;Yu-Jing Tian;Jia Cao(Department of Ophthalmology,Wuxi Children's Hospital,Wuxi 214023,Jiangsu Province,China;Department of Ophthalmology,Wuxi People's Hospital,Wuxi 214023,Jiangsu Province,China)

机构地区：[1]中国江苏省无锡市儿童医院眼科,214023 [2]中国江苏省无锡市人民医院眼科,214023

出　　处：《国际眼科杂志》2018年第6期1004-1009,共6页International Eye Science

基　　金：无锡市卫生局青年项目(No.Q201405)~~

摘　　要：目的:利用DNA探针杂交技术,结合显色探针技术建立一种新型、高灵敏的免疫检测体系,用于早期先天性白内障的筛查。方法:选取3个常染色体显性遗传的先天性白内障家系中患者14例,取静脉血并提取mR NA,建立CRYAB的捕获探针及显色探针。利用DNA探针,通过碱基配对原则形成三明治结构(捕获探针-DNA探针-显色探针)检测入选者的血样。1家系6例患者静脉血利用酶联免疫吸附测定(ELISA)法检测αB-晶状体蛋白。结果:最佳条件下,双特异探针技术可检测到最低浓度的先天性白内障晶状体蛋白的突变基因,各突变位点检测率为99.5%～99.7%;ELISA法检测样本αB-晶状体蛋白上调,阳性率为85.9%。双特异探针技术敏感性更高,检测位点更多,ELISA法仅局限于蛋白检测水平,精确性不高。结论:双特异探针检测技术操作简单,灵敏度高,可重复性高,经济实惠,在临床上用于产前诊断、优生优育具有重要的应用价值。AIM:To screen congenital cataract in the early stage using dual-specific probe technique composed of DNA hybridization and chromogenic probe.METHODS:The mRNA was isolated from venous blood of 14 congenital cataract patients with autosomal dominant inheritance in 3 chromosomes.Based on genetic mutats of CRYAB gene,DNA probe,capture probe and signal probe were designed.The sandwich structures containing capture probe,DNA probe and signal probe was used to detect genetic mutants in 8 samples from one family;and ELISA was used to detect the contents fluctuation of Crystallin alpha B.RESULTS:The dual-specific probe technique detected the minimum genetic mutation of Crystallin alpha B in congenital cataract samples,various mutations detection rate was between 99.5%and 99.7%.In ELISA detection,serum Crystallin alpha B level increased,and detection rate was 85.9%.Compared with ELISA assay,this novel assay was more sensitive.CONCLUSION:The novel dual-specific probe method is quite convenient for detection of genetic mutants of congenital cataract;and for its high sensitivity and repeatability,it is of great potential in clinically prenatal diagnosis,and it might play significant roles in eugenic and superior nurture.

关 键 词：双特异探针技术 先天性白内障 晶状体蛋白基因 

分 类 号：R776.1[医药卫生—眼科]