Programmable DNA nanotube mediated molecular machine and exosome nucleic acid detection method thereof

Through a molecular machine based on programmable DNA nanotube-mediated, combining the synergistic effect of strand displacement amplification (SDA) and DNA nanotubes, ultra-sensitive detection of exosomal nucleic acid miR-193b is achieved, solving the problem of distinguishing Alzheimer's disease patients from normal people in the prior art, and improving the sensitivity and specificity of the detection.

CN119709949APending Publication Date: 2025-03-28SHANDONG RES INST OF TUMOUR PREVENTION TREATMENT
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Patent Information

Application Number
CN202411995418.0
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2024-12-31
Publication Date
2025-03-28

AI Technical Summary

Technical Problem

The prior art is difficult to accurately distinguish Alzheimer's disease patients from normal people, and the existing nanomaterial-based detection platforms have problems such as low signal amplification efficiency, high detection limits and insufficient specificity.

Method used

Using a molecular machine based on programmable DNA nanotube mediated, ultra-sensitive detection of exosomal nucleic acid miR-193b is achieved through the synergistic effect of strand displacement amplification (SDA) and DNA nanotubes. The method includes complementary pairing of the target exosome nucleic acid miR-193b with the template sequence Probe A, catalyzed extension and cleavage using Klenow polymerase to generate Probe B, which mixes with blocked DNA nanotubes to open the ribozymes to form activated DNA nanotubes, and rolling the substrate chain Probe C with ferrocene Fc signal tags on the surface of the gold electrode to generate an amplified electrochemical signal.

Benefits of technology

It realizes ultra-sensitive detection of exosomal nucleic acid miR-193b, with a detection limit of 5.7fM, with high specificity and rapid response capabilities, and is suitable for early disease diagnosis and treatment monitoring.

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Abstract

The invention discloses a molecular machine based on programmable DNA nanotube mediation and a detection method of exosome nucleic acid of the molecular machine, and the preparation method of the molecular machine based on programmable DNA nanotube mediation is characterized by comprising the following steps: (1) complementarily pairing target exosome nucleic acid miR-193b and a template sequence Probe A to obtain a product extension chain; (2) carrying out catalytic extension and cutting on an extension chain by using Klenow polymerase, i.e., carrying out multiple rounds of polymerization and cutting on the product in the step a to generate a large amount of Probe B for subsequent reaction; and (3) enabling S1, S2, S3-DNAzyme, S4 and S5 to react in a TM buffer solution at 37 DEG C for 1 hour, so as to prepare the blocked molecular machine based on the programmable DNA nanotube mediation. The technology realizes high-sensitivity and high-selectivity nucleic acid detection, and has important clinical diagnostic value.
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