离子强度对乙型脑炎病毒基因传感器检测系统的影响

Effects of Ionic Strength on Japanese Encephalitis Virus Gene Sensor Detection System

目的探讨磷酸盐缓冲液(PBS)中Na+浓度值对乙型脑炎病毒(JEV)基因传感器检测系统的影响。方法漏声表面波(LSAW)基因传感器表面固定终浓度为1.0μmol/L的乙型脑炎病毒探针,然后分别在pH7.6的10mmol/LPBS缓冲液(含Na+浓度值0.1～0.6mol/L)中和终浓度为1.0μmol/L的JEV靶序列杂交,分别记录相位变化及反应所需时间。结果杂交反应引起的相位变化是先上升后趋于缓和的趋势,以0.3mol/LNa+浓度为分界线;杂交平衡时间同相位变化一致,当PBS中Na+浓度为0.3mol/L,杂交反应的平衡时间为33min。结论随着Na+浓度的升高,杂交反应引起的相位变化呈典型的饱和曲线趋势,PBS中Na+浓度0.3mol/L,杂交反应平衡时间33min,是LSAW-JEV基因传感器杂交的最适反应条件。

OBJECTIVE To investigate the effects of Na ionic concentrations in phosphate batfered saline （PBS） on hybridization efficiency with Japanese encephalitis virus （JEV） gene sensor detection system. METHODS The concentration of 1.0μmol/L of JEV probe was immobilized on the surface of leaky surface acoustic were （LSAW） gene sensor, then 1.0μmol/L JEV target sequence was hybridized with the probe while pH 7.6 10mmol/L PBS buffer contained different concentrations Na ionic varied from 0.1 to 0.6mol/L,and then the changes in phase and equilibrium time of reaction were recorded and analyzed. RESULTS Phase increased firstly and then trended gently when Na ionic concentrations from 0.1 to 0.6mol/L, and 0.3mol/L was the watershed. And equilibrium time for hybridization change tendency coincided with phase, equilibrium time of hybridization reaction was 33 min at 0.3mol/L Na ionic concentrations. CONCLUSIONS The phase change caused by hybridization reaction is a saturation curve in accordance with the increase in Na ionic concentration. Therefore, 0.3mol/L Na ionic concentrations of pH 7.6 10mmol/L PBS buffer and 33 rain equilibrium time of hybridization reaction are optimized work conditions of LSAW-JEV gene sensor.