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目的建立一种基于电化学生物传感器的快速检测埃博拉病毒核酸的方法。方法利用一步热变性法自组装成带有固定探针的DNA四面体后,通过Au-S键固定至金电极表面制备成新型核酸探针。与合成的埃博拉病毒核酸序列特异性结合后引入Bio-ss DNA检测序列,通过生物素和链霉亲和素的特异性结合作用引入avidinHRP放大信号,利用计时电流法进行检测。结果该传感器可特异性识别和定量检测人工合成的埃博拉病毒的核酸序列,通过实验条件优化,测定核酸的浓度范围在1.0×10~(-9)~5.0×10~(-6)mol/L呈良好的线性关系,检测下限为5.2×10~(-10)mol/L。结论所构建的DNA四面体探针修饰的电化学生物传感器实现了对埃博拉病毒核酸的灵敏、特异检测。
Objective To establish a rapid electrochemical detection method for Ebola virus based on electrochemical biosensor. Methods The DNA tetrahedra with immobilized probes were self-assembled by one-step thermal denaturation. Then, a new type of nucleic acid probe was prepared by Au-S bond to the surface of gold electrode. Bio-ss DNA was introduced into the specific sequence of the synthetic Ebola virus to detect the sequence. Avidin HRP was amplified by the specific binding of biotin and streptavidin, and the signal was detected by chronoamperometry. Results The sensor could specifically detect and quantitatively detect the nucleic acid sequence of synthetic Ebola virus. The optimal concentration of nucleic acid was 1.0 × 10 -9 ~ 5.0 × 10 -6 mol / L showed a good linear relationship, the detection limit of 5.2 × 10 ~ (-10) mol / L. Conclusion The constructed DNA biosensor with electrochemical tetrahedron probe has realized the sensitive and specific detection of Ebola virus nucleic acid.