Objective   To investigate the effects of different concentrations of target sequence of Japanese encephalitis virus (JEV) on the hybridization efficiency with leaky surface acoustic wave (LSAW) gene sensor detection system.     Methods   After 0.5 μmol/L JEV probe was firstly immobilized on the surface of LSAW gene sensor

JEV target sequence at different concentrations of 0.05

0.1

0.5

1.0

1.5

and 2.0 μmol/L was hybridized with the probe in pH 7.6 10 mmol/L PBS buffer (containing Na ionic concentrations of 0.3 mol/L)

and then the changes of phase and equilibrium time of reaction were recorded and analyzed.     Results   Phase variation with JEV probe was gradually increased firstly and then in a steady trend when the concentrations of target sequence were changed from 0.05 to 2.0 μmol/L. Concentration of 1.0 μmol/L was determined as the watershed. No evident change was observed in equilibrium time for hybridization except for the concentration of 0.05 μmol/L. Equilibrium time of hybridization reaction was 33 min when the concentration of at JEV target sequence was 1.0 μmol/L.    Conclusion   The phase variation caused by hybridization reaction is saturation curve in accordance with the increase of target sequence concentration. Therefore

JEV target sequence at a concentration of 1.0 μmol/L and equilibrium time for 33 min in hybridization reaction are optimized work conditions for LSAW gene sensor.