缓冲液pH值对LSAW-bisPNA传感器检测系统的影响  

Effect of pH value of buffer on LSAW-bisPNA gene sensor detection system

作　　者：张烨[1] 府伟灵[1] 陈鸣[1] 王云霞[1] 曹亮[2] 丁毅[2] 齐永志[1] 

机构地区：[1]第三军医大学西南医院检验科,重庆400038 [2]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》2007年第12期1161-1163,共3页Journal of Third Military Medical University

基　　金：国家"863"计划重大专项(2002AARZ2023);国家自然科学基金(30400107);全军"十一五"课题(06G073);重庆市科技攻关项目(8327)~~

摘　　要：目的探讨PBS缓冲液pH值对LSAW-bisPNA基因传感器检测系统的影响。方法LSAW-bisPNA基因传感器表面固定终浓度为1.0μmol/L的bis-PNA探针,然后分别在pH5.8～8.0的PBS缓冲液中和相应的靶序列杂交,记录相位变化及反应所需时间。结果在不同pH值缓冲液中,bis-PNA与靶序列反应引起的相位变化有明显差异(P<0·01),pH6.6与pH6.8时传感器响应信号最大,但两者反应所达到的平衡时间有明显差异(P<0·05),pH值6.6时检测所需时间更短。结论选择pH6.6作为bis-PNA和靶序列dsDNA反应的最适pH值,在该pH值条件下传感器响应信号大,反应达平衡时间短。Objective To investigate the effects of pH values of PBS buffer on hybridization efficiency with leaky surface acoustic wave （LSAW） bis-peptide nucleic acid （bisPNA） gene sensor detection. Methods Firstly 1.0 μmol/L bis-PNA probe was immobilized on the surface of LSAW-bisPNA gene sensor, then hybridized with corresponding human papilloma virus （HPV） target sequence when pH values of PBS varied from 5.8 to 8.0, and the changes of phase and equilibrium time of reaction were recorded and analyzed. Results The changes of phase varied significantly in different pH values of buffer. And the signal reached highest when pH was 6.6 and 6.8. However, the equilibrium time for hybridization had significant difference between pH 6.6 and pH 6.8 （P 〈 0. 05 ）, the time for pH 6.6 was shorter. Conclusion pH 6.6 is the most suitable for the hybridization of bisPNA and target dsDNA when the signal is utmost and the equilibrium time is short.

关 键 词：PH值 肽核酸 漏声表面波 基因传感器 杂交 

分 类 号：R312[医药卫生—基础医学] R313