Applications of gene-chip for the detection of mutations in cTnI gene associated with FHCM  

家族性肥厚型心肌病相关cTnI基因突变检测芯片的应用(英文)

作　　者：张元颖[1,3] 郭会时[1] 何农跃[1,2] 谷宇[1] 杨迪[2] 万文辉[2] 张寄南[2] 

机构地区：[1]东南大学生物电子学国家重点实验室,南京210096 [2]南京医科大学第一附属医院心血管病研究所,南京210029 [3]江苏省肿瘤防治研究所

出　　处：《Journal of Southeast University(English Edition)》2005年第3期330-335,共6页东南大学学报（英文版）

基　　金：The National Natural Science Foundation of China(No.60071001);China Postdoctoral Science Foundation(No.2002);Trans-Century Training Programme Foundation for the Talents by theState Education Commission(No.[2004]527);the Foundation of the 135?Key Laboratory of Jiangsu Province(No.SK200205);the HighTechnology Research Plan of Jiangsu Province(No.BG2003033,BG2001010).

摘　　要：Mutations in cardiac troponin I （cTnI） gene were assessed based on gene-chip technology.Special probes were designed to fabricate the low-density gene-chip,which could detect the mutations in exons 3,5,7,and 8 of the cTnI gene simultaneously.For each exon,two oligonucleotide sequences labeled with fluorescein at the 5＇-end were designed,one （oligonucleotide Ⅰ） simulating the wild type and the other （oligonucleotide Ⅱ） simulating the mutant.Oligonucleotides Ⅰ and Ⅱ were mixed together to simulate the heterozygote.After optimizing the hybridization protocols,the fabricated gene-chip could detect the mutations in the exons of the cTnI gene with relative high sensitivity and specificity.The fully complementary probe gave a fluorescent signal almost 50% stronger than that of the one-base mismatched one,which is in accordance with the result from a theoretical estimate. An applicable special gene-chip is available to investigate and diagnose familial hypertrophic cardiomyopathy （FHCM） after further improvement.基于基因芯片技术对心肌钙蛋白(cTnI)基因上的突变进行了分析.针对外显子上的突变特征设计了特异性探针,制备了可以同时检测cTnI基因上第3,5,7,8外显子突变的低密度基因芯片.对每一个外显子,设计了2条5′端标记荧光的寡核苷酸链,一条模拟野生型序列,另一条模拟突变型序列,将二者混合起来模拟杂合子序列.经过实验条件的优化,制作的芯片可以灵敏、特异地检测cTnI基因外显子上的突变.结果表明,该芯片检测突变正配错配区别明显,荧光强度比值符合理论估计(正中碱基错配的信号强度是完全正配信号强度的50%左右).经进一步对芯片优化后,该芯片有望在家族性肥厚型心肌病的研究和诊断中得到应用.

关 键 词：hypertrophic cardiomyopathy(HCM) GENE-CHIP MUTATION HYBRIDIZATION 

分 类 号：Q81[生物学—生物工程]