低毒纳米晶电化学发光与信号放大用于乳腺癌细胞和相关microRNA的高灵敏检测与成像分析

Breast cancer is one of the women’s most common malignancy, it is of crucial significance for early diagnosis of breast cancer to develop new, rapid, highly sensitive and selective method for detecting breast cancer cells and related miRNA. In this project, we intend to detect breast cancer cells and miRNA by using highly sensitive electrochemiluminescence of low-toxic nanocrystals. We would prepare novel, low-toxic nanocrystals and nanocomposites with excellent ECL properties, and establish new techniques to enhance ECL of nanocrystals. Based on specific biological recognition, combining nano-amplification and surface resonance-enhancing ECL effects between gold nanoparticles and nanocrystals, we would develop novel nanocrystal biological probe with amplified ECL signal which can specifically recognize cancer cells and miRNA. We would build nano-amplified biosensing interface by using graphene oxide (GO), silver nano-cluster and PAMAM materials etc., and develop new highly sensitive ECL biosensing strategy to detect cancer cells and miRNA by using multiple enzyme-aided cyclic amplification signal techniques with sensitive nanocrystal ECL biological probe. We would develop new nanocrystal ECL imaging analysis techniques, and achieve in situ ECL imaging and visual analysis of breast cancer cells and related miRNA, which would provide more accurate detection technology and evidence for early diagnosis of breast cancer.

乳腺癌是女性最常见的恶性肿瘤之一，发展快速、高灵敏、高选择性的检测癌细胞及相关miRNA的新方法对乳腺癌早期诊断具有极其重要的意义。本项目以乳腺癌细胞及相关miRNA为检测对象，以高灵敏的纳米晶电化学发光（ECL）为检测技术，拟开发制备新型、低毒、高ECL性能的纳米晶及其复合物，建立增强纳米晶ECL的新技术；基于特异性的生物识别反应，结合纳米放大效应、金属与纳米晶之间的表面共振增强ECL效应，研制新型、具有放大信号的、特异性识别癌细胞或miRNA的纳米晶ECL生物探针；利用氧化石墨烯、银纳米簇、PAMAM等材料构建具有纳米放大效应的生物传感界面，将多种酶循环放大信号的技术与纳米晶ECL生物探针相结合，发展高灵敏检测乳腺癌细胞及miRNA的ECL生物传感新策略；建立纳米晶ECL成像分析新技术，实现乳腺癌细胞及miRNA的ECL原位成像和可视化分析，为乳腺癌早期诊断提供更为准确的检测技术和依据

发展快速、高灵敏、高选择性检测肿瘤细胞及相关miRNA的新方法对有效解决肿瘤早期诊断这一临床迫切的实际问题，具有极其重要的意义。该项目开发了系列新型、低毒、高效ECL性能的纳米复合物，并对其结构表征、电化学发光性能及ECL信号提高进行了研究。制备了新型的Ru(bpy)32+-SiO2纳米复合材料、PDDA-GO-石墨烯量子点、Ag(I)离子增强ECL信号的CdSe量子点、Au NPs/ZnSe QDs复合物、AgNCs、碳量子点等ECL信号材料，研究了ECL性能及提高ECL信号的方法技术。制备了系列新型的纳米晶ECL探针，发展了高灵敏的ECL生物传感方法及信号放大技术。构建了新型CdSe/ZnS量子点纳米簇ECL信号探针，研制了高灵敏ECL生物传感器，实现了对乳腺癌细胞高选择性ECL检测。利用Ru@SiO2 ECL发光剂，通过目标循环放大技术研制了新型ECL生物传感器。研制了基于循环放大反应和DNA walker技术的纳米晶ECL传感器，实现了对目标miRNA的高灵敏ECL检测。构建了新型Ag(I)离子增强CdSe量子点ECL信号的生物传感平台，用于miRNA等双目标的“开、关”检测。利用新型PAMAM-Au复合材料负载大量CdSe量子点，构建了信号放大的ECL生物探针，实现了对DNA的ECL检测。构建了高灵敏检测乳腺癌相关miRNA的ECL生物传感新方法。利用PDDA-GO-石墨烯量子点ECL探针，构建了高灵敏ECL 生物传感器。利用DNA水凝胶和Walker DNA多重信号放大生物传感技术，实现了乳腺癌相关miRNA灵敏检测。利用DNA纳米管放大信号技术，研制了电化学发光多功能生物传感器，实现了双目标高灵敏检测。利用DNAzyme扩增策略与DNA-walker诱导构象转换相结合，开发了通用的ECL多功能生物传感平台。利用CdSe@ZnS和CdTe量子点信号探针，实现了双目标MicroRNA的同时检测。基于DNA超支化杂交链式反应结合DNA walker扩增技术，构建了树枝状多功能检测平台，实现了癌细胞的荧光成像。