基于电化学适体信标技术的HIV-1 p24抗原快速检测新方法研究

HIV-1 p24抗原检测能显著缩短HIV感染检测窗口期，但酶免法操作繁琐，发光法设备要求高，均难以满足现场快速筛查要求。电化学传感器以其简便、快速、准确、价廉的特点，成为 p24抗原检测技术发展新方向。作为传统电化学传感器识别元件的抗体在环境中易失活，且普通材料修饰电极电传导性能有限，鉴于这些缺点，我们在前期免疫传感芯片研究工作基础上，拟用SELEX技术筛选出亲和力强、特异性高、性质稳定的HIV-1 p24抗原核酸适体，构建胭脂红酸电化学活性-非活性开关适体信标（CAs-AMB）作为识别元件，配合新型纳米材料-石墨烯复合物作为修饰的工作电极，并通过磁性富集方法固定CAs-AMB，构建一种新型的HIV-1 p24抗原检测电化学传感器，并深入解析该传感器液相响应模式，完成其检测性能的临床评价，为HIV-1 p24抗原快速现场筛查提供一种简便、灵敏、准确的新方法。

p24 antigen, the HIV-1 capsid protein, is of considerable diagnostic interest due to its detectable possibility several days earlier than HIV antibodies screening following HIV exposure. But the complexity and poor sensitivity of conventional ELISA method and the expensive equipment requirement for chemiluminescence method both fail to meet rapid screening of HIV-1 p24 antigen...Electrochemical biosensor has become a very competitive detection method for HIV infection markers because of excellent analytical properties,such as convenience,speed,accuracy and low cost.Traditional electrochemical biosensors have shortcomings,include the inactivation of antibodies as the recognition components in compound environment and the poor performance of ordinary material modified electrodes in electrosignal transduction. These unsatisfied situation draw us to develop a new electrochemical biosensor consisted of carminic acids made electrochemically active-inactive switching HIV-1 p24 aptamer molecular beacon(CAs-AMB) as the recognize element, and the new work electrode modified with a newly developed nanomaterial-graphene comlplex. We schedule to concentrate the CAs-AMB onto electrode to enhance the sensitivity through the superparamagnetic Fe3O4 nanoparticles,a method we have developed during the past research experiments...We will also examine the response mechanism using the mathematics model on this novel HIV-1 p24 biosensor and its diagnostic characteristics in clinic evaluation compared with the chemiluminesence method. We hope this newly developed HIV-1 p24 CAs-AMB biosensor could be used in the rapid sreening of HIV-1 early infection.