摘要
目的 初步构建适于临床基因检测的新方法即压电基因传感器阵列。方法 制作 2× 5型基因传感器阵列 ,在阵列各检测池的银膜上通过巯基修饰法固定合成大肠杆菌肠毒素基因探针 ,而后与各靶序列进行杂交 ,以自制集成电路及电脑软件对频率数据进行采集与分析。结果 12h内 ,气相对照检测池频率变化± 2Hz ,液相对照检测池频率变化± 4Hz ;基因探针固定后频率下降 ( 4 8± 5 )Hz ;与完全配对的靶序列杂交 ,0 .5h达到稳定状态 ,并使频率下降 ( 4 3± 5 )Hz ;与有 1～ 2个碱基错配的序列杂交 ,频率下降仅分别为 ( 2 1± 5 )、( 7± 5 )Hz ,与 3个碱基错配的序列及不相关序列杂交 ,频率无明显变化。结论 初步构建的基因传感器阵列有很好的稳定性和特异性 ,敏感性尚待进行一步提高。
Objective To construct the piezoelectric sensor arrays for gene identification in clinical work. Methods Gene sensor arrays of 2×5 were made, gene probes were immobilized on Ag surface, then hybridized with 8 kinds of oligonucleotides, data were collected and analyzed by self made integrated circuit and computer soft ware. Results In 12 hours, frequency change of gas and liquid controls were less than ±2 Hz and ±4 Hz , respectively. Frequency cut down (48±5)Hz after probe immobilization and (43±5)Hz after a full matched hybridization for 0.5 hours. Frequency cut down only (21±5)Hz and (7±5)Hz when hybridized with the sequences of 1 and 2 bases mismatching. No change was found following hybridization with target sequences of 3 bases mismatching and non related target sequences. Conclusion The constructed gene sensor arrays is good in stability and specificity, but need to be improved in sensitivity.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2001年第7期810-812,共3页
Journal of Third Military Medical University
基金
国家自然科学基金资助项目 ( 39870 831)
国家科技部攻关项目 ( 95 A2 3 0 4 0 4 )
