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目的 探讨自行研发的压电基因传感器对链置换扩增 (SDA)的实时检测。方法 以巨细胞病毒 (HCMV)为检测对象 ,建立成熟的 SDA反应系统 ;在自行研发的压电基因传感器检测仪上对巨细胞病毒 SDA反应进行实时检测。结果 SDA反应可引起压电基因传感器的频率持续下降 ;在一定范围内核酸杂交所引起的频率下降幅度与加入的靶序列浓度呈正相关 ;加样时温差等因素对早期频率变化曲线的影响不可忽视 ;实验温度是影响SDA压电基因传感器检测灵敏度的重要因素。结论 自行研发的压电基因传感器可成功实现对 SDA反应的实时检测 ;该系统可直接检测基因组 DNA并可广泛应用于病原微生物的临床检测。
Objective To investigate the real-time detection of strand displacement amplification (SDA) by self-developed piezoelectric gene sensor. Methods Cytomegalovirus (HCMV) was used as a detection target to establish a mature SDA reaction system. Real-time detection of cytomegalovirus SDA was performed on a self-developed piezoelectric gene sensor detector. Results The frequency of piezoelectric gene sensor decreased continually due to SDA reaction. The decrease of frequency caused by nucleic acid hybridization was positively correlated with the concentration of target sequence added in a certain range. The influence of temperature difference on the frequency curve of early stage could not be neglected The experimental temperature is an important factor that affects the sensitivity of the SDA piezoelectric gene sensor. Conclusion The self-developed piezoelectric gene sensor can achieve real-time detection of SDA reaction. The system can directly detect genomic DNA and can be widely used in the clinical detection of pathogenic microorganisms.