A method for separating and enriching circulating tumor dna

Abstract

The invention provides a method for separating and enriching circulating tumor DNA, comprising the following steps: step 10) centrifuging the plasma sample to obtain a first supernatant; step 20) centrifuging the first supernatant to obtain a second supernatant supernatant; step 30) centrifuging the second supernatant to obtain a third supernatant; step 40) removing fibrin in the third supernatant to obtain a fourth supernatant; step 50) removing the fourth supernatant exosomes in the supernatant to obtain a fifth supernatant. The separation and enrichment method of circulating tumor DNA provided by the present invention can remove cells and large fragments after the first centrifugation treatment, remove blood platelets after the second centrifugation treatment, and remove large cysts after the third centrifugation treatment The fourth time can remove fibrin, the fifth time can remove exosomes, and the content of circulating tumor DNA in the final fifth supernatant is higher than that obtained by the existing method.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for separating and enriching circulating tumor DNA. Background technique [0002] Circulating tumor DNA (corresponding to English: circulating tumor DNA, abbreviated in the text: ctDNA) refers to the continuous flow in the human blood circulation system carrying certain characteristics (including mutations, deletions, insertions, rearrangements, copy number abnormalities, methylation, etc.) from DNA fragments of the tumor genome. As a non-invasive cancer marker, ctDNA has important development prospects in the fields of in vitro diagnosis, medication and prognosis. [0003] ctDNA is derived from tumor cells and widely exists in many tumor types. However, the concentration of ctDNA is low, highly fragmented, and the proportion of mutation sites that can be detected is very low, so separation and enrichment are particularly important. The process of extracting c...

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Problems solved by technology

However, the concentration of ctDNA is low, highly fragmented, and the proportion of mutation sites that can be detected is very low, so separation and enrichment are particularly important

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