差分脉冲和方波伏安法研究亲和素-生物素固载ssDNA的生物传感器  

Study on DNA biosensor based on immobilization ssDNA by avidin-biotin system with differential pulse voltammetry and square wave voltammetry

作　　者：杜晓燕[1] 郑丽[1] 王茂清[1] 公维磊[1] 

机构地区：[1]哈尔滨医科大学公共卫生学院,哈尔滨150081

出　　处：《中国卫生检验杂志》2007年第3期425-427,共3页Chinese Journal of Health Laboratory Technology

基　　金：黑龙江省自然科学基金资助项目(D2006-35)

摘　　要：目的：建立一种新型的DNA检测技术，能快速、灵敏的识别目的DNA。方法：生物素标记的ssDNA通过生物素-亲和素体系固载到Pt电极上，制作了DNA传感器，[Co（phen）3]^3作指示剂，用差分脉冲伏安法（DPV）和方波伏安法（SWV）对此传感器进行研究。结果：在pH6．8—7．0，[Co（phen）3]^3＋浓度80μmol／L条件下，杂交前后指示剂峰电流的差值与溶液中互补DNA的浓度有良好的线性关系，DPV法测量起始子的线性范围是8．0×10^-9 ～2．8×10^-8 mol／L，终止子的线性范围是7．2×10^-9 ～7．2×10^-8mol／L；SWV法测量起始子的线性范围是8．5×10^-9 ～3．5×10^-8mol／L，终止子的线性范围是7．2×10^-9 ～9．0×10^-8 mol／L。两种检测方法均能很好的识别样品中的DNA片断。结论：该方法测量灵敏度高，能很好地识别转基因食品中的特异DNA序列。Objective ：To develop a method for rapid and sensitive detection of target DNA. Methods：Biotin -labelled ssDNA was immobilized on the platinum electrode surface by means of avidin - biotin system. DPV and SWV were employed to transduce the hybridization event by using [ Co（phen） 3 ] ^3＋ as an electroactive indicator. Results： Under pH 6. 8 - 7.0 and [ Co（phen） 3 ] ^3＋ concentration of 80 μmol/L, an excellent linearity between the differences of the peak currents before and after hybridization and the concentration of complementary DNA was achieved. The depend linearity detected by DPV of the promoter and terminator respectivly was 8.0 × 10^ -9 - 2. 8 × 10 ^-8 mol/L and 7. 2 × 10^-9 - 7. 2 × 10^-8 mol/L. The depend linearity detected by SWV of the promoter and terminator respectivly was 8.5 × 10^ -9 - 3. 5 × 10^-8 mol/L and 7. 2 × 10 ^-9 - 9.0 × 10^ -8 mol/L. It was applied com- mendably to detect transgenic soybean power. Conclusion ：The DNA biosensor can be used to detect target DNA sensitively.

关 键 词：差分脉冲伏安法 方波伏安法 生物素-亲和素 DNA传感器 

分 类 号：O657.1[理学—分析化学]