基于Cu-MOF-还原氧化石墨烯的电化学免疫传感器检测膀胱癌二级预防生物标志物的研究

Bladder cancer (BC) is a common type of malignant neoplasms in the urinary system; it has some clinical characteristics such as an increasing incidence, the morbidity concealment, high degree of malignancy and a high rate of recurrence after treatment. The bladder tumor antigen (BTA) in human urine sample plays an important role in the early detection, diagnosis and treatment of BC. Therefore, developing a low cost, highly sensitive and rapid method for the detection of BTA is quite crucial for early detection and diagnosis of BC. Compared with the traditional method, electrochemical immunosensor has a unique advantage in the detection of BTA. In this work, some highly sensitive electrochemical immunosensors for detection of BTA will be developed by using newly prepared and functional modified silica (SiO2)-modified copper-metal organic framework (Cu-MOF), reductive graphene-tetraethylenepentamine (rGO-TEPA) and their composite materials modified screen-printed electrode (SPE), respectively. The proposed electrochemical immunosensor in this work will show satisfied performance such as simple fabrication, low detection limits, wide linear range, short analysis time, high stability and selectivity in the detection of BTA. Furthermore, the proposed electrochemical immunosensor will be employed to test real urine samples of patients with newly diagnosed BC and will require satisfactory results compared with other traditional method such as ELISA and IRMA. The proposed electrochemical immunosensor will also provide a new method and a new idea for the detection of other urine biomarkers of BC and other tumor biomarkers.

膀胱癌是泌尿系统最常见的恶性肿瘤之一，具有发病率高、发病隐匿、恶性程度高以及治疗后高复发等特点。尿液中膀胱肿瘤抗原在早期发现、早期诊断以及早期治疗膀胱肿瘤中发挥着重要的作用，因此实现其快速、特异、灵敏、高效的检测意义重大。同传统检测方法比，电化学免疫传感器在膀胱肿瘤生物标志物检测中具有独特的优势。本项目拟采用新制备并功能化修饰的二氧化硅包被铜-MOF材料、还原氧化石墨烯-四亚乙基戊胺及两者复合材料作为电极修饰材料修饰丝网印刷电极，然后在其表面固定膀胱肿瘤抗原的抗体，并对不同电极修饰材料、修饰方法以及抗体固定方法等进行全面、系统评价与优化，筛选出最佳组合方案制备电化学免疫传感器检测尿液中膀胱肿瘤抗原。本项目构建的电化学免疫传感器具有灵敏度高、选择性好、应用范围广等诸多优点，可以为多种膀胱癌尿液生物标志物以及其他肿瘤生物标志物的检测提供新方法和新思路，具有十分广阔的应用前景。

膀胱癌是泌尿系统常见恶性肿瘤之一，具有发病率高、发病隐匿、恶性程度高以及治疗后高复发等特点。尿液中膀胱癌生物标志物在早发现、早诊断及早治疗膀胱癌中发挥至关重要的作用，因此实现膀胱癌生物标志物的快速、特异、灵敏、准确检测具有重要意义。本研究采用水热法和搅拌法成功合成Cu-MOFs@SiO2和ZIF8两种MOFs材料，并将两者分别与还原氧化石墨烯-四亚乙基戊胺（rGO-TEPA）和贵金属纳米粒子（AuNPs，AgNPs）相结合，成功合成rGO-TEPA@Cu-MOFs@SiO2@Ag、rGO-TEPA@Au-ZIF8两种复合材料，将不同材料的优势充分结合，并采用多种表征技术对合成的材料进行表征。在评估不同材料性能的基础上，采用一次性的丝网印刷电极（SPE）作为基底电极，顺利构建两种电化学免疫传感器用于膀胱癌生物标志物核基质蛋白22（NMP22）的检测。（1）基于rGO-TEPA@Cu-MOFs@SiO2@Ag的电化学免疫传感器，该传感器的线性范围为0.0001到1000 ng/mL，检测限为33.33 fg/mL；（2）基于rGO-TEPA@Au-ZIF8的电化学免疫传感器，该传感器的线性范围为0.01到1000 ng/mL，检测限为3.3 pg/mL。此外，两种传感器具有良好的选择性、稳定性、可重复性和准确性，能够用于实际样品的检测。本项目构建的电化学免疫传感器具有灵敏度高、选择性好、应用范围广等优点，可以为多种膀胱癌生物标志物以及其他肿瘤标志物的检测提供新方法和新思路。

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