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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.
Based on the gene chip technology, the mutation of cardiac troponin (cTnI) gene was analyzed.A specific probe was designed according to the mutation characteristics of exon, Exon mutations in the low-density gene chip.For each exon, designed two 5 ’end labeled fluorescent oligonucleotide chain, a simulated wild-type sequence, another analog mutant sequence, the two mixed To simulate the heterozygous sequence.After the optimization of the experimental conditions, the prepared chip can detect the mutation of the cTnI gene exon in a sensitive and specific manner.The results show that the mutation detection positive mutation has a significant difference, and the fluorescence intensity ratio accords with the theoretical estimation (Median base mismatch signal strength is completely positive with signal strength of about 50%.) After further chip optimization, the chip is expected to be used in the research and diagnosis of familial hypertrophic cardiomyopathy.