Kit for extracting DNA from human feces sample, extraction method and application

Abstract

The invention relates to the technical field of medical biology, in particular to a kit for extracting DNA from a human feces sample and an extraction method. The kit comprises a sample preservation solution, a cell lysis solution, a DNA precipitant, a DNA detergent, a DNA extracting solution and a DNA eluent, the sample preserving solution comprises the following components: a trihydroxymethyl aminomethane hydrochloric acid buffer, sodium chloride, ammonium sulfate and ethylenediamine tetraacetic acid, the cell lysis solution comprises the following components: guanidinium isothiocyanate, a chelating agent, dithiothreitol, TritonX-100 and a pH regulator; and the DNA extracting solution comprises guanidine hydrochloride. When the extracting solution is used for extracting DNA from an fecessample, the situation that a DNA adsorption column is blocked in the extracting process is not likely to happen, and the extracting success rate is high. By adopting the extraction method disclosed by the invention, the concentration and purity of the DNA sample solution can be greatly improved, and the extracted DNA sample solution can be well used for methylation detection.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a kit, an extraction method and an application for extracting DNA from human feces samples. Background technique [0002] With the development of molecular biology, liquid biopsy technology based on the detection of exosomes, CTC (circulating tumor cells), and cfDNA (free DNA (deoxyribonucleic acid)) has shown unparalleled advantages over traditional detection methods. The advantages of liquid biopsy technology are high sensitivity, high specificity, and non / low invasiveness. [0003] For diseases of the digestive system, the DNA released when the gastrointestinal epithelium, especially the epithelial cells of the lesions undergo apoptosis or death, can more effectively reflect the specific conditions of the digestive system of the subject. However, normal human feces are an extremely mixed mixture, including water and solids, among which, solids include dead bacteria, fat, ...

AI Technical Summary

Problems solved by technology

However, normal human feces are an extremely mixed mixture, including water and solids, among which, solids include dead bacteria, fat, protein, inorganic salts and undigested residual food and solid components in digestive juices, while DNA is dispersed in Among the mixtures, how to find and extract qualified DNA from so many mixtures for later detection is a difficult problem in the current non-invasive detection technology of feces

[0004] At present, the kits used for fecal DNA extraction have technical problems of complex composition and low extraction success rate

For the stool DNA extraction method, there are technical problems that the extraction purity is not high, or the concentration of DNA extracted from the same amount of stool is not high, and it cannot be applied to different shapes of stool

At the same time, the existing technology also has the technical problem that the extracted fecal DNA cannot be used for methylation detection.

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