The HIV-DNA detection platform based on Au nanoparticle DNA probe and ExoⅢ-assisted signal amplification
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摘要:
设计了一种金纳米粒子(AuNP)和核酸外切酶Ⅲ(ExoⅢ)辅助靶标循环信号放大的新型纳米探针用于HIV-DNA的检测.该纳米探针由AuNP和捕获探针p1和信号探针p2构成.在靶标HIV-DNA不存在的情况下,此时纳米探针抗ExoⅢ水解,信号探针p2与AuNP之间的距离较近产生荧光共振能量转移,体系的荧光基本被猝灭.但是,在靶标存在的情况下,靶标与信号探针p2杂交,将p1置换下来,此时与靶标杂交的信号探针p2双链体能在ExoⅢ的作用下发生水解,使荧光团释放,体系荧光增强.同时,靶标也被释放并与AuNP上的探针p2作用,以驱动下一个反应,达到靶标循环目的,因此这种方式检测时间较短且稳定性高.该方法的线性范围为100 pmol/L~50 nmol/L,检测限为38.68 pmol/L(信噪比S/N=3).该方法具有良好的抗干扰性,可用于复杂生物样品检测,回收率在98.72%~106.72%之间.
A novel nanoprobe with Au nanoparticles(AuNP)and ExoⅢassisted target circulating signal amplification was designed for HIV-DNA detection.The nanoprobe is composed of AuNP,capture probe p1 and signal probe p2.In the absence of the target HIV-DNA,the nanoprobe is resistant to ExoⅢhydrolysis at this time,and the distance between the signal probe p2 and AuNPs is close to produce fluorescence resonance energy transfer,and the fluorescence of the system is basically quenched.However,in the presence of a target,the target hybridizes to the signal probe p2 and replaces p1.At this time,the signal probe p2 duplex that hybridizes with the target can hydrolyze under the action of ExoⅢ,the fluorophore is released,and then the system fluorescence is enhanced.At the same time,the target is also released and interacts with the probe p2 on AuNPs to drive the next reaction in order to achieve the goal of target circulation,so this method has a short detection time and high stability.Therefore,this method has higher detection sensitivity.The linear range of this method is 100 pmol/L-50 nmol/L,and the detection limit is 38.68 pmol/L(S/N=3).With good anti-interference ability,the method can be used for the detection of complex biological samples with a recovery rate between 98.72%-106.72%.
作者:
田建袅 李晴 梁雪花 谭莉 陆江南 王鑫
Tian Jianniao;Li Qing;Liang Xuehua;Tan Li;Lu Jiangnan;Wang Xing(Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources(Ministry of Education of China,School of Chemistry and Pharmaceutical Science,Guangxi Normal University,Guilin 541004,China))
机构地区:
广西师范大学化学与药学学院
出处:
《betway官方app 学报：自然科学版》 CAS 北大核心 2021年第2期54-62,F0002,共10页
基金:
国家自然科学基金(22064003 21465007 21565007)。
关键词:
HIV DNA 金纳米粒子 核酸外切酶Ⅲ 信号放大
HIV-DNA Au nanoparticle ExoⅢ signal amplification
分类号:
O657.3 [理学—分析化学]
