乙型脑炎病毒靶序列浓度对LSAW基因传感器检测系统的影响  

Concentrations of target sequence of Japanese encephalitis virus in LSAW gene sensor detection system

作　　者：徐清华[1] 陈鸣[1] 陈伟[1] 府伟灵[2] 王云霞[2] 丁毅[3] 施建峰[3] 曹亮[3] 汪广杰[2] 

机构地区：[1]第三军医大学大坪医院野战外科研究所检验科,重庆400042 [2]第三军医大学西南医院检验科,重庆400038 [3]中国电子科技集团公司第二十六研究所第五研究室,重庆400060

出　　处：《第三军医大学学报》2009年第9期765-767,共3页Journal of Third Military Medical University

基　　金：国家高技术研究发展计划(863计划)(2007AA02Z416);国家自然科学基金(30400107)~~

摘　　要：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 encephalitis virus （JEV） on detection system. Methods sensor, JEV target sequence To investigate the effects of different concentrations of target sequence of Japanese the hybridization efficiency with leaky surface acoustic wave （LSAW） gene sensor After 0.5μmol/L JEV probe was firstly immobilized on the surface of LSAW gene 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 Ixmol/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 33min in hybridization reaction are optimized work conditions for LSAW gene sensor.

关 键 词：漏声表面波 基因传感器 相位 乙型脑炎病毒 杂交 

分 类 号：R318.04[医药卫生—生物医学工程] R373.31[医药卫生—基础医学]