基于稀土上转换材料的高危型HPV-DNA荧光探针的构建及其细胞成像研究

Cervical cancer has become one of the serious malignant tumors causing women to death and threating their health. Studies have shown that persistent or repetitive infection of high-risk human papilloma virus (high-risk HPV) is the bottom cause in the development of cervical cancer. So fast, reliable and effective HPV detection is very important for screening and prevention of cervical cancer. DNA is the core substance of HPV and basis of virus replication, which is the most direct, specific and sensitive index of HPV infection. Early and accurate detection of DNA sequences of high-risk HPV is extremely important for the disease diagnosis and treatment. The current DNA detection methods suffer from high cost and they are prone to false positive results in clinical application. This project aims to construct novel rare erath up-conversion nanoparticles (Re-UCNPs) for highly sensitive high-risk HPV-DNA detection based on FRET (fluorescence resonance energy transfer). Re-UCNPs and fluorescent dyes will act as energy donor and energy acceptor in the FRET process, respectively. This study will not only extend the applications of Re-UCNPs in the biological applications, but also enrich HPV-DNA detection. Meanwhile, it has the reference value for diagnosis and treatment of cervical cancer.

宫颈癌是严重危害妇女健康和生存的恶性肿瘤之一。研究表明高危型人乳头瘤病毒（HPV）持续性或反复性感染已成为宫颈癌发生的根本原因，快速、可靠和有效的HPV检测对筛查和预防宫颈癌非常重要。HPV-DNA是HPV病毒的核心物质和病毒复制的基础，是HPV感染最直接、特异性强和灵敏性高的指标，因此及早和准确的检测引起该疾病的DNA序列对于高危型HPV患者的诊断和治疗具有重要意义。针对目前临床中HPV-DNA检测技术成本高、误差大、易出现假阳性结果等问题，本项目提出以荧光共振能量转移原理为介导，构建以稀土上转换发光材料为能量给体，荧光染料为能量受体的DNA荧光探针，实现高危型HPV-DNA的痕量检测。本项目的实施将拓展上转换发光材料在生物标志方面的应用，丰富HPV-DNA的检测手段，对宫颈癌的早期诊断和治疗具有参考价值。

工作设计合成了多个新型荧光探针，筛选出2个用于HPV-DNA的研究，同时对金属离子荧光探针以及在细胞内的应用展开研究。筛选出基于罗丹明B为发光团的Hg2+荧光探针，在其它金属离子和阴离子浓度很高的情况下也不会对Hg2+的分析检测产生干扰，这一特征使其很好的满足了生物学和环境监测方面的要求；另外项目还实现了单一荧光探针对Mg2+、Al3+的同时检测，以及对Cu2+的比率分析，进一步用于细胞内成像检测令人满意。