石英谐振传感器液相稳定平台构建及检测HCMV的实验研究

Design of a stable system of quartz oscillation sensor in liquid phase and the test of HCMV

目的 建立石英谐振压电基因传感器检测系统液相稳定平台并应用该检测系统实时检测链置换扩增(Stranddisplacement amplification,SDA)反应。方法①观察以金属夹具式和粘胶式两种检测池连接方式构成的传感器在液相中的频率稳定性;②以巨细胞病毒(Human cytomegalovirus,HCMV)为检测对象,建立SDA反应系统;③传感器检测系统对HCMVSDA反应进行实时检测。结果①新型金属夹具式可调节及可换式传感器检测池可实现传感器在液相中的频率稳定性;②SDA反应可引起压电基因传感器的频率持续下降;③在一定范围内核酸杂交所引起的频率下降幅度随加入的靶序列浓度提高而增大。结论 成功构建压电基因传感器液相检测稳定平台并实现对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 design the stable system of piezoelectric gene sensor in liquid phase and detect strand displacement amplification(SDA) reaction with the piezoelectric gene sensor. Methods The frequency stability of pools with metal clamping and glue connecting way in liquid phase was observed. SDA reaction system was established using human cytomegalovirus( HCMV) as the detecting subject. Real time monitoring of HCMV SDA reaction was performed by using the piezoelectric gene sensor system. Results A new sensor-testing pool with the metal clamping of the adjustable and exchangeable style was able to solve the problem of the frequency stability in liquid phase. SDA could lead to continuous reduction of frequency in piezoelectric gene sensor. The degree of frequency reduction resulted from gene reaction was positively correlated with the consistency of the target gene in a certain range. Conclusion We have designed the system of piezoelectric gene sensor in liquid phase and conducted real time monitoring of the detection of SDA successfully. The system can be used for the direct detection of gene DNA as well as in the clinical detection of pathogenic microbes.