Extraction reagent for exosome extraction as well as application and extraction method thereof

Abstract

The invention discloses an extraction reagent for exosome extraction. The extraction reagent is prepared from the following components: dithiothreitol, potassium chloride, potassium citrate, sodium formate, serum albumin, glycine and guanidinium thiocyanate, and has a pH value of 3.0-5.5. The extraction method comprises the following steps: a, centrifuging a body fluid sample through a centrifugal machine to obtain supernate; b, adding the extraction reagent into the supernate obtained in the step a, reacting for 2 hours, and centrifuging through the centrifugal machine; and c, suspending and re-dissolving the exosome deposit obtained in the step b with PBS to obtain exosome. The exosome extraction is simple in operation and short in time consumption, the operation procedure is simplified, the cost is saved, and clinical routinization of body fluid biopsy becomes possible.

Description

technical field [0001] The invention relates to the field of clinical medicine, in particular to an extraction reagent suitable for exosome extraction and its application and extraction method. Background technique [0002] Exosomes are rapidly extracted from body fluids (urine, blood, saliva, ascites, ascites, etc.) and nucleic acid molecules (DNA, RNA, and miRNA), which play important roles in intercellular substance and signal transduction in vivo. Because these nucleic acids are encapsulated and protected, they have high stability and are not easy to degrade. They are ideal new biomarkers for tumor diagnosis and prognosis monitoring. Early diagnosis, drug monitoring, and prognosis judgment of cancer. [0003] In recent years, with the deepening of people's research and understanding of exosomes, exosome detection, as a new liquid biopsy hotspot technology, has been widely used by many clinical research institutions for non-invasive diagnosis, treatment and monitoring of...

AI Technical Summary

Problems solved by technology

[0004] At present, the standard for exosome extraction is still ultracentrifugation; the method is very complicated and time-consuming. Taking the extraction of exosomes in urine as an example, at least 200 ml of urine is required each time, and ultracentrifugation (120000g / min) It takes more than 20 hours to obtain sufficient exosomes. At the same time, the equipment requirements of the ultracentrifuge and the training of the operating procedures greatly increase the extraction cost, and it is impossible to achieve clinical routine application.

Due to the lack of relevant exosome extraction reagents in my country, the research on exosomes in China basically relies on the cumbersome ultracentrifugation and imported extraction kits, which seriously hinders the development of exosome research and clinical application in China.

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