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Reagent for preserving nucleic acid of feces exfoliated cells and its preparation method and application

A technology for preserving reagents and cell nucleic acids, which is applied in the field of fecal exfoliated cell nucleic acid preservation reagents and its preparation, which can solve the problems of adverse effects of detection, no removal of peculiar smell, etc., and achieve the effects of increasing stability, avoiding interference, and simplifying downstream operation steps

Active Publication Date: 2021-01-22
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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  • Reagent for preserving nucleic acid of feces exfoliated cells and its preparation method and application
  • Reagent for preserving nucleic acid of feces exfoliated cells and its preparation method and application
  • Reagent for preserving nucleic acid of feces exfoliated cells and its preparation method and application

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Experimental program
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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 a nucleic acid preservation reagent of feces exfoliated cells and its preparation method and application. The nucleic acid preservation reagent of feces exfoliated cells of the present invention includes nonionic surfactants, anionic surfactants, osmotic pressure regulators, cell membrane stabilizers, chelating agents Mixture, pH buffer and water, also contains deodorant. The preservation reagent of the present invention can effectively lyse human exfoliated cells in feces, and does not contain toxic components such as guanidine thiocyanate and guanidine isothiocyanate, thereby providing a method for directly extracting and detecting feces genome from feces supernatant , can effectively avoid the interference of fecal impurities, and there is no need to precipitate the fecal sample again before testing.

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 Patents(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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