Preparation of circulating tumor free DNA (deoxyribonucleic acid) standard product for simulating plasma matrix

Abstract

The invention provides preparation of a circulating tumor free DNA (deoxyribonucleic acid) standard product for a simulating plasma matrix. The preparation comprises the following steps of A, purchasing a natural cell line carrying gene mutation, or editing the gene to obtain a cell line carrying the gene mutation; B, cracking a wild cell line and a gene mutation cell line, and obtaining fragmented DNA by a nucleosome enzyme digestion or ultrasonic intercepting method; mixing the fragmented DNA of the wild cell line and one or multiple fragmented DNA of the gene mutation cell line according toa ratio, so that the mixture has the required gene mutation points and mutation frequency; D, preparing simulating plasma; E, feeding the mixture in step D into the simulating plasma, fully and uniformly mixing, and verifying the property. The preparation has the advantages that a standard product of a ctDNA sample approximate to a natural sample of the human body is provided, and a standard reference substance is provided for a circulating tumor free DNA detection platform, development of detection reagents, and evaluation on laboratory detection ability.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the preparation of a circulating tumor free DNA standard product simulating a plasma matrix. Background technique [0002] At present, tumor has become the disease with the highest morbidity and mortality in the world, which seriously threatens human health and survival. Therefore, early diagnosis is crucial to improve patient prognosis. There are various gene mutations in the process of tumorigenesis. Detection of mutation types in patients not only helps to understand the mechanism of tumorigenesis, but also can be used as molecular markers for tumor susceptibility assessment and diagnosis. At present, fresh tissues and paraffin-embedded tissues are mostly used in the detection of tumor gene mutations in clinical practice. The detection results can guide the treatment of patients, but there are still some problems: (1) Local samples cannot reflect tumor heterogeneity. (2) It is n...

AI Technical Summary

Problems solved by technology

At present, fresh tissues and paraffin-embedded tissues are mostly used in the detection of tumor gene mutations in clinical practice. The detection results can guide the treatment of patients, but there are still some problems: (1) Local samples cannot reflect tumor heterogeneity

(3) Organizational detection cannot implement dynamic monitoring

The software, hardware and reagent development of the above-mentioned technical methods, as well as the evaluation of laboratory detection capabilities, require standard substances to measure, so circulating tumor cell-free DNA standards that simulate plasma stroma need to be developed urgently

Images