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目的探讨乙型脑炎病毒(Japanese encephalitis virus,JEV)靶序列浓度值对漏声表面波(LSAW)基因传感器检测系统的影响。方法LSAW基因传感器表面固定终浓度为0.5μmol/L的JEV探针,然后用pH7.6的10mmol/L PBS缓冲液(含Na+0.3mol/L)中和终浓度为0.05、0.1、0.5、1.0、1.5、2.0μmol/L的JEV靶序列杂交,分别记录相位变化及反应所需时间。结果与JEV探针反应引起的相位变化是先缓慢上升后趋于缓和的趋势,以1.0μmol/L靶序列浓度为分界线;杂交平衡时间上除0.05μmol/L的时间短以外,其他的未见明显变化趋势,当靶序列浓度为1.0μmol/L,杂交反应平衡时间为33min。结论随着靶序列浓度的升高,杂交反应引起的相位变化呈典型的饱和曲线趋势;靶序列浓度为1.0μmol/L、杂交反应的平衡时间为33min,是LSAW基因传感器杂交的最适反应条件。
Objective To investigate the effect of JEV (target encephalitis virus) target sequence on detection system of LSAW gene sensor. Methods The JEV probe with a final concentration of 0.5 μmol / L was immobilized on the surface of LSAW gene sensor, then neutralized with 10 mmol / L PBS buffer (containing Na + 0.3 mol / L) at a pH value of 7.6 for a final concentration of 0.05, 0.1, 0.5, 1.0, 1.5, 2.0μmol / L JEV target sequence hybridization were recorded phase changes and the reaction time required. Results The phase change induced by JEV probe reaction firstly tended to be alleviated after slowly rising. The concentration of 1.0 μmol / L target sequence was the dividing line. When the time of hybridization was shorter than 0.05 μmol / L, See obvious trends, when the target sequence concentration of 1.0μmol / L, hybridization reaction equilibrium time of 33min. Conclusion As the concentration of target sequence increases, the phase change caused by the hybridization reaction shows a typical trend of saturation curve. The target sequence concentration is 1.0μmol / L and the equilibrium time of hybridization is 33min, which is the optimum reaction condition for LSAW gene sensor hybridization .