CN105779597A - 一种基于分裂式核酸适配体双金属纳米探针的单分子识别方法 - Google Patents

一种基于分裂式核酸适配体双金属纳米探针的单分子识别方法 Download PDF

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CN105779597A
CN105779597A CN201610191362.2A CN201610191362A CN105779597A CN 105779597 A CN105779597 A CN 105779597A CN 201610191362 A CN201610191362 A CN 201610191362A CN 105779597 A CN105779597 A CN 105779597A
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noble metal
target
aptamer
recognition method
probe
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洪昕
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Dalian University of Technology
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Abstract

本发明属于生物医学及医药研究中的单分子影像与识别技术领域。其特征是目标靶标的核酸适配体被一分为二形成两个片段，每个片段在其分割端标记上具有表面等离子体共振效应的贵金属纳米粒子，形成两类贵金属粒子标记的核酸适配体分子探针；以两个分别来自上述两类中的探针在绑定到同一靶标分子而复原时所产生的稳定的表面等离子体共振耦合信号作为靶标的特异性单分子识别信息。本发明所能达到的效果是实现在众多单个贵金属标记物存在的噪声背景中识别出目标靶标，具有特异性目标识别能力，解决目前使用金属标记物特异性识别能力差的问题。

Description

一种基于分裂式核酸适配体双金属纳米探针的单分子识别方法
技术领域
本发明属于生物医学及医药研究中的分子影像与识别技术领域，涉及到基于金属探针的分子影像技术，特别涉及到一种基于金属纳米探针的高特异性光学识别方法。
背景技术
影像学在临床诊断中占有重要的地位，传统的影像学对疾病的诊断是以病理改变为基础的，发现时往往是疾病的中末期。分子影像技术是在分子水平检测病变，从而真正达到早期发现，早期诊断、早期干预/治疗的目的。分子的尺寸很小，若想对它们进行光学显影，需要借助于标记物。荧光物质是普遍采用的一种标记物，它的探测灵敏度高、技术成熟。但是荧光物质也存在着光致漂白或“眨眼”等缺点而使观测时间受限，因此使得其它可替代标记物质日益受到重视，贵金属纳米粒子就是其中之一。例如金纳米粒子具有高亮度、可无限时间观测、生物兼容性好，更为突出的是其具有表面等离子体共振特性(SPR)。在其SPR波长处展现出强烈的吸收，为热疗和光学成像提供了可操作空间。以上这些优点使得其作为生物分子探针的标记物在生医相关的基础研究和重大疾病的早期临床诊断等领域极为期待。目前亟待解决的瓶颈问题之一是如何进行目标靶标的特异性识别，这是因为被金属粒子标记后的分子探针进入样品之后，都可以通过金属粒子进行显影。如何从这些影像中中识别出哪些是绑定到了目标靶标上的粒子、哪些是未发生绑定反应的粒子是进行开展诊断的关键。
发明内容
本发明的目的是提出一种基于分裂式核酸适配体双金属探针的分子识别方法，能够通过纳米级金属标记物进行特异性生物反应识别，为基于金属纳米标记物的分子影像技术提供一种特异性分子识别方法。
本发明的技术解决方案是，将目标靶标的核酸适配体分割为两段，在每一段的切割端连接上一个纳米级贵金属粒子，构成一对带有金属标记物的核酸适配体分子探针；当这样形式的一对分子探针绑定到同一个目标靶标时，它们将两个贵金属纳米粒子拉到一起；两个靠近的粒子由于表面等离子体共振耦合，在其缝隙处产生强烈的增强电场，其吸收可比单粒子增强2～数百倍；以此信号作为特异性分子绑定反应的识别信息。
所述的标记的贵金属纳米粒子是具有表面等离子体共振效应。
所述的用于标记的贵金属纳米粒子具有纳米结构。
所述的用于标记的两个贵金属纳米粒子的纳米结构相同或不同。
本发明所能达到的效果是实现在众多单个金属纳米标记物存在的噪声背景中识别出目标靶标，具有特异性目标识别能力，突破目前使用金属纳米标记物特异性识别能力差的瓶颈。
附图说明
附图1是基于分裂式核酸适配体双金属探针的分子识别方法示意图。
1是目标靶标；
2是目标靶标的核酸适配体；
3是将2一分为二的其中之一片段，并在分割端标记上贵金属粒子；
4是将2一分为二的其中另一片段，并在分割端标记上贵金属粒子；
5是3、4的两种分子探针共同绑定到目标靶标时，将两个金属粒子拉在一起，在双金属粒子的缝隙处产生增强信号，以此信号作为单分子靶标的特异性识别信号。
具体实施方式
例如图1所示，目标靶标为一段单链DNA，其核酸适配体为一段与目标靶标具有互补碱基序列的DNA分子；将该核酸适配体一分为二，分别获得片段3和片段4；分别对片段3和片段4连接金纳米粒子构成一对标记了金纳米粒子的分子探针；将如此获得的两种探针等量注入到待测样品中，两个分裂的核酸适配体片段在探测到同一靶标分子并与其绑定复原后，在双粒子的间隙处产生表面等离子体共振耦合增强信号，用于特异性识别单分子靶标。

Claims (5)

1.一种基于分裂式核酸适配体双金属纳米探针的单分子识别方法，其特征是将待测靶标的一段核酸适配体分裂为二段，在每一段的分裂端分别标记上贵金属纳米粒子，形成两类分子探针；当一对分别来自于上述两个种类的探针绑定到同一个靶标，在复原后将两个贵金属纳米粒子拉拢靠近，在该对粒子的间隙处产生的表面等离子体共振耦合增强信号作为单分子靶标的特异性识别信息。
2.如权利要求1所述的一种基于分裂式核酸适配体双金属纳米探针的单分子识别方法，其特征是标记的贵金属纳米粒子是具有表面等离子体共振效应。
3.如权利要求1或2所述的一种基于分裂式核酸适配体双金属纳米探针的单分子识别方法，其特征是用于标记的贵金属纳米粒子具有纳米结构。
4.如权利要求1或2所述的一种基于分裂式核酸适配体双金属纳米探针的单分子识别方法，其特征是用于标记的两个贵金属纳米粒子的纳米结构相同或不同。
5.如权利要求3所述的一种基于分裂式核酸适配体双金属纳米探针的单分子识别方法，其特征是用于标记的两个贵金属纳米粒子的纳米结构相同或不同。
CN201610191362.2A 2016-03-28 2016-03-28 一种基于分裂式核酸适配体双金属纳米探针的单分子识别方法 Pending CN105779597A (zh)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106370868A (zh) * 2016-09-23 2017-02-01 中国科学院重庆绿色智能技术研究院 基于核酸适配体信号放大策略的检测微囊藻毒素的spr传感器及其制备方法和应用

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* Cited by examiner, † Cited by third party
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HUILIN TAO等: "Highly Sensitive Resonance Scattering Detection of DNA Hybridization Using Aptamer-Modified Gold Nanopaticle as Catalyst", 《PLASMONICS》 *
JAMES J. STORHOFF等: "One-Pot Colorimetric Differentiation of Polynucleotides with Single Base Imperfections Using Gold Nanoparticle Probes", 《J. AM. CHEM. SOC.》 *
JAMES J. STORHOFF等: "Programmed Materials Synthesis with DNA", 《CHEM. REV.》 *
R. BUICULESCU等: "The optimization of oligonucleotide conjugation onto gold nanoparticles for biodetection", 《R. BUICULESCU等》 *
李玉佩: "基于核酸适配体与金纳米粒子探针检测凝血酶的共振光散射分析", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 *
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106370868A (zh) * 2016-09-23 2017-02-01 中国科学院重庆绿色智能技术研究院 基于核酸适配体信号放大策略的检测微囊藻毒素的spr传感器及其制备方法和应用

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