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| 基于量子点荧光共振能量转移(FRET)效应的裂开型CD20核酸适配体探针用于非霍奇金淋巴瘤的检测研究* |
| Detection of NHL cells based on split CD20 aptamer and QDs-mediated FRET effect * |
| 投稿时间：2018-12-06 修订日期：2018-12-12 |
| DOI： |
| 中文关键词: 核酸适配体 CD20 量子点 FRET 非霍奇金淋巴瘤 |
| 英文关键词: Aptamer CD20 quantum dot FRET NHL |
| 基金项目:陕西省重点研发项目(2017-SF-280)；西安市科技计划项目(SF1510(4)) |
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| 中文摘要: |
| 目的：通过CD20核酸适配体CE4-1分裂为两段并分别标记量子点和荧光受体,基于靶细胞可诱导上述两段裂开片段形成特定识别构型拉近荧光供受体对进而导致FRET效应的原理,成功构建了一种新型的NHL激活式检测荧光探针。方法：将CD20核酸适配体CE4-1进行裂解,结合量子点和Cy5荧光供受体队之间的FRET效应,利用流式细胞术,对裂开型核酸适配体的裂开位置、比例、浓度和反应时间进行优化,并在最优条件下评估该检测体系对CD20+细胞的检测特异性。结果：在CE4-1-1a和CE4-1-2b浓度比例为1:5、CE4-1-1a浓度为4nM、孵育时间为50min时,染料标记在裂开位点处等一系列条件下,该探针显示出对靶肿瘤细胞最强的亲和力和FRET信号激活性能。同时,该探针可有效保持对CD20+细胞的高特异性。结论：基于量子点荧光共振能量转移(FRET)效应的裂开型CD20核酸适配体探针体系在检测CD20+细胞方面表现出了极低的背景信号,同时有着较好的FRET信号值,有望实现CD20+ NHL细胞的高灵敏激活式检测。 |
| 英文摘要: |
| Objective: Based on the principle that these two pecies could be induced by target to form a certain recognition conformation and then make the donor-acceptor pair close to initiate FRET，CD20 aptamer termed as CE4-1 was splited and a novel activatable fluorescent detection probe for CD20+ cells was successfully constructed．Methods: CD20 aptamer CE4-1 was splited and modified with QD or Cy5. Based on the FRET effect and flow cytometry, the split site, ratio, concentration and incubation time of detection system were optimized. Further, the detection specificity of system was evaluated. Results: It was demonstrated that the system was composed of CE4-1-1a and CE4-1-2b with the mole ratio of 1:5, the CE4-1-1a concentration of 4 nM and incubated with cells in 50min, it exhibited relatively strong FRET signal to target tumor cells. Further, this splited aptamer system showed recognization specificity to CD20+ cells. Conclusions: The CD20 splited aptamer system based on FRET for the detection of NHL exhibits weak background signal and relatively strong FRET signal, which presents a promising method for detection of CD20+ NHL cells. |
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