Colocalization trigger chain type hybridization reaction based aptamer sensor for detecting adenosine

An aptamer sensor and hybridization reaction technology, applied in the field of adenosine aptamer sensor detection of adenosine, to achieve good reproducibility and precision, eliminate interference, and reduce non-specific reactions

Active Publication Date: 2014-11-19
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the problem that the existing adenosine analysis technology cannot realize the direct detection of adenosine in biological samples, the purpose of the present invention is to provide an aptamer sensor based on colocalization to trigger chain hybridization reac...

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  • Colocalization trigger chain type hybridization reaction based aptamer sensor for detecting adenosine
  • Colocalization trigger chain type hybridization reaction based aptamer sensor for detecting adenosine
  • Colocalization trigger chain type hybridization reaction based aptamer sensor for detecting adenosine

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Embodiment 1

[0056] Example 1 Constructing an aptamer sensor based on colocalization-triggered chain hybridization for the detection of adenosine

[0057] (1) Instruments and reagents

[0058] (1) Instrument device

[0059] Fluorescence spectrophotometer (model F-2500, Hitachi, Japan); electric thermostatic incubator (DNP-9052, Shanghai Jinghong Experimental Equipment Co., Ltd.); dry thermostat (K30, Hangzhou Aosheng Instrument Co., Ltd.); vortex mixing Instrument (VDRTEX-5, Haimen Qilin Bell Instrument Manufacturing Co., Ltd.); balance (H-101, Shanghai Kangle Optoelectronic Instrument Co., Ltd.); electronic balance (ME model, Mettler-Toledo Instrument Co., Ltd.).

[0060] (2) Reagents

[0061] The nucleic acid probes used in this experiment were synthesized and purified by Yingwei Jieji (Shanghai) Trading Co., Ltd., as follows:

[0062] Biotinylated aptamer strand-1 (Bio-Apt-1) is:

[0063] 5'-biotin-TTTTTTTTTTTT GTG CAA GTA CCA TT TAG ACC ACC TGG GGG AGT AT-3' (the nucleotide sequenc...

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Abstract

The invention discloses a colocalization trigger chain type hybridization reaction based aptamer sensor for detecting adenosine. The colocalization trigger chain type hybridization reaction based aptamer sensor consists of an aptamer chain 1, an aptamer chain 2 and a hairpin probe which are biotinylated and a magnetic nano-sphere modified by streptavidin. The aptamer sensor is used for detecting adenosine, and a detection method comprises the following steps: (1) activation of a streptavidin modified magnetic sphere; (2) fixation of the biotinylated aptamer chain 1; (3) colocalization of adenosine-induced double aptamers; (4) colocalization trigger chain type hybridization reaction; (5) addition of a dye SYBR Green I and fluorescence detection; (6) calculation: calculating the concentration of adenosine in a to-be-detected object containing adenosine according to a measured fluorescence value. The aptamer sensor disclosed by the invention can be used for reducing the non-specific reaction of conventional chain type hybridization reaction by virtue of the colocalization-induced hybridization reaction, has the advantages of high detection sensitivity and less sample usage without enzyme amplification, and can achieve the analysis and detection of low-concentration adenosine.

Description

technical field [0001] The invention relates to the construction of an adenosine aptamer sensor based on colocalization triggered chain hybridization reaction to detect adenosine. Background technique [0002] Adenosine is an important biopharmaceutical small molecule, which participates in the regulation of various life activities of the body, and has important physiological and pharmacological effects, such as relaxing blood vessels, contracting smooth muscles, and regulating the nervous system and immune system. In addition, the detection of adenosine in biological samples can be used for disease diagnosis, monitoring of disease course and early diagnosis of cancer. Therefore, the quantitative detection of adenosine, especially the detection of adenosine in biological samples, has important biological significance in clinical research, disease diagnosis and prevention, and early diagnosis of cancer. [0003] Although a large number of adenosine aptamer sensing systems ha...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/682C12Q2525/205C12Q2525/301C12Q2563/107
Inventor 王磊姜玮冯春景富波
Owner SHANDONG UNIV
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