Exosome detection method

Abstract

The invention relates to an exosome detection method which comprises the following steps: activating a carboxylated magnetic bead, generating an aptamer probe, covalently binding the aptamer probe to the surface of the carboxylated magnetic bead to obtain a magnetic bead-aptamer probe, adding an auxiliary agent into the magnetic bead-aptamer probe, binding to obtain a magnetic bead-aptamer probe-auxiliary agent, and detecting the exosome by using the magnetic bead-aptamer probe-auxiliary agent. The method comprises the following steps: combining an exosome with a magnetic bead-aptamer probe-adjuvant, carrying out centrifugation to obtain a magnetic bead-aptamer-exosome and a probe-adjuvant, carrying out a chromogenic reaction on a color developing agent and the probe-adjuvant to generate a fluorescent substance, and finally detecting the fluorescent substance. The method has the advantages that the reaction steps are simple, and raw materials are not easily influenced by environmental factors.

Description

technical field [0001] The present application relates to the technical field of biological detection, in particular to an exosome detection method. Background technique [0002] Exosomes are vesicular bodies secreted by living cells with a size of approximately 30–150 nm. Exosomes contain a variety of important substances, including proteins, RNA, lipids, etc., which carry rich molecular information on primitive cells and play an important role in various physiological and pathological processes. Tumor-associated exosomes are an important mechanism for regulating the occurrence and development of tumors, and are considered as potential markers for early tumor diagnosis. Therefore, it is crucial to develop sensitive and specific assays for tumor exosomes. [0003] Currently, many methods are used to detect exosomes, such as dynamic light scattering (DLS), nanoparticle tracker (NTA), surface plasmon resonance (SPR), nanoflow cytometry, etc. The above detection methods need...

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Problems solved by technology

The above detection methods need to rely on expensive large-scale instruments, and the detection steps are complicated

[0004] In addition, exosome detection methods also include enzyme-linked immunoassay and western blot analysis. Although both enzyme-linked immunoassay and western blot analysis do not need to rely on expensive large-scale instruments and have high sensitivity, both There are still problems, that is, the antibody protein is easily deformed by the influence of pH value, temperature and other environmental factors, and the operation steps of the whole detection process are relatively complicated. As far as the enzyme-linked immunoassay is concerned, it needs to go through 8 times of washing work, and the reaction time of each step Up to 30min

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