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Excrement exfoliated cell nucleic acid preservation reagent as well as preparation method and application thereof

A technology for preserving reagents and cell nucleic acids, applied in the field of fecal exfoliated cell nucleic acid preservation reagents and their preparation, can solve problems such as adverse effects of detection and no odor removal, and achieve the effects of increasing stability, avoiding interference, and simplifying downstream operation steps.

Active Publication Date: 2020-05-26
WUHAN AIMISEN LIFE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are also some reagents and methods for preserving stool samples at room temperature. For example, in Chinese patent application 20151067125.8, the main function of the preservation reagents is to maintain the cell shape and prevent cell rupture. In the next step of detection, the fecal impurities in the sediment are likely to have an adverse effect on the detection

Method used

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  • Excrement exfoliated cell nucleic acid preservation reagent as well as preparation method and application thereof
  • Excrement exfoliated cell nucleic acid preservation reagent as well as preparation method and application thereof
  • Excrement exfoliated cell nucleic acid preservation reagent as well as preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1 feces preservation reagent formula

[0031] Feces preservation reagent 1: 0.5% Triton X-100, 0.1% SDS, 50mM EDTA, 50mM Tris, 150mM sodium chloride, 2% mannitol, pH 8.0;

[0032] Feces Preservation Reagent 2: 0.5% Triton X-100, 0.1% SDS, 50mM EDTA, 50mM Tris, 150mM Sodium Chloride, 2% Mannitol, 0.1% Lauryl Methacrylate, 5% Ethanol, 0.1% Peppermint Essential Oil, 0.25 % Jasmine essential oil, pH8.0;

[0033] Feces preservation reagent 3: 0.5% Triton X-100, 0.1% SDS, 50mM EDTA, 50mM Tris, 150mM sodium chloride, pH 8.0;

[0034] Feces preservation reagent 4: 0.5% Triton X-100, 0.1% SDS, 50mM EDTA, 50mM Tris, 150mM sodium chloride, 2% mannitol, 1% deodorant A (Lanjing deodorant deodorant, purchased from Jinan Delan Chemical Co., Ltd., its main ingredients are active ingredients extracted from plants such as yucca, ginkgo leaves, tea polyphenols, grape seeds, Lauraceae plants, eucalyptus oil, pine oil), pH 8.0.

Embodiment 2

[0035] Embodiment 2 preserves reagent preparation method

[0036] Preparation method of stool preservation reagent 1: Measure 5mL Triton X-100, weigh 1g SDS powder, 6.06g Tris powder, 14.61g EDTA powder, 8.77g sodium chloride powder, 20g mannitol powder, add purified water to make up to 1L , adjust the pH value to 8.0 with hydrochloric acid.

[0037] Preparation method of stool preservation reagent 2: Measure 5mL Triton X-100, weigh 1g SDS powder, 6.06g Tris powder, 14.61g EDTA powder, 8.77g sodium chloride powder, 20g mannitol powder, 1g lauryl methacrylate, Add 50mL of ethanol, 1g of peppermint essential oil, 2.5g of jasmine essential oil, add purified water to make up to 1L, and adjust the pH value to 8.0 with hydrochloric acid.

[0038] Preparation method of stool preservation reagent 3: Measure 5mL Triton X-100, weigh 1g SDS powder, 6.06g Tris powder, 14.61g EDTA powder, 8.77g sodium chloride powder, add purified water to 1L, adjust pH with hydrochloric acid The value i...

Embodiment 3

[0040] The extraction of embodiment 3 stool total DNA

[0041] The kit used is QIAamp DNA Stool Mini kit (QIAGEN), and the specific operation steps can be found in the kit manual.

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Abstract

The invention provides an excrement exfoliated cell nucleic acid preservation reagent as well as a preparation method and an application thereof. The excrement exfoliated cell nucleic acid preservation reagent comprises a nonionic surfactant, an anionic surfactant, an osmotic pressure regulator, a cell membrane stabilizer, a chelating agent, a pH buffer solution, water and a deodorant. The preservation reagent disclosed by the invention can be used for effectively cracking human exfoliated cells in excrement, and does not contain guanidinium thiocyanate, guanidinium isothiocyanate and other toxic components, so that a method for directly extracting and detecting the excrement genome from an excrement supernatant is provided, the interference of the excrement impurities can be effectively avoided, and the excrement sample does not need to be subjected to precipitation treatment before the detection.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a reagent for preserving nucleic acid of feces exfoliated cells and its preparation method and application. Background technique [0002] The human body will have a certain amount of exfoliated cells entering the feces every day. Studies have shown that these exfoliated cells contain cancerous signals, so fecal samples are ideal samples for non-invasive detection of certain cancers such as digestive tract cancers, and have great potential in the field of cancer detection in the future. It has great application prospects. In 2014, Exact sciences in the United States launched a multitarget stool DNA test kit for non-invasive detection of colorectal cancer. [0003] The composition of feces is very complex, which contains animal DNA, plant DNA and a wide variety of bacterial DNA, as well as PCR inhibitors such as polysaccharides, bile salts, humic substances...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12Q1/6806C12Q1/6886C12Q1/6883
CPCC12Q1/24
Inventor 张良禄吴志诚董兰兰袁奥兰甘甜李婷婷
Owner WUHAN AIMISEN LIFE TECH CO LTD
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