|Table of Contents|

[1] Zhang Yuanying, Guo Huishi, He Nongyue, et al. Applications of gene-chip for the detection of mutationsin cTnI gene associated with FHCM [J]. Journal of Southeast University (English Edition), 2005, 21 (3): 330-335. [doi:10.3969/j.issn.1003-7985.2005.03.017]
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Applications of gene-chip for the detection of mutationsin 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

Journal of Southeast University (English Edition)[ISSN:1003-7985/CN:32-1325/N]

Volumn:
21
Issue:
2005 3
Page:
330-335
Research Field:
Biological Science and Medical Engineering
Publishing date:
2005-09-30

Info

Title:
Applications of gene-chip for the detection of mutationsin 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
Author(s):
Zhang Yuanying1, 3, Guo Huishi1, He Nongyue1, 2, Gu Yu1, Yang Di2, Wan Wenhui2, Zhang Jinan2
1 State Key Laboratory of Bioelectronics, Southeast University, Nanjing 210096, China
2 Research Institute of Cardiovascular Disease, First Affiliated Hospital, Nanjing Medical University, Nanjing 210029, China
3Jiangsu Institute of Cancer Research, Nanjing 210009, China
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
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, 南京 210009
Keywords:
hypertrophic cardiomyopathy(HCM);  gene-chip;  mutation;  hybridization
肥大型心肌病;  基因芯片;  突变;  杂交
PACS:
Q81
DOI:
10.3969/j.issn.1003-7985.2005.03.017
Abstract:
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.

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Memo

Memo:
Biographies: Zhang Yuanying(1978—), female, master;He Nongyue(corresponding author), male, doctor, professor, nyhe1958@163.com.
Last Update: 2005-09-20