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Excrement preserving fluid and applications thereof

A technology for preserving liquid and feces, applied in the field of microbial detection, can solve problems such as high cost, impact on preservation quality, and limitation, and achieve the effects of ensuring accuracy, good preservation effect, and safe operation

Active Publication Date: 2018-05-04
CAPITALBIO GENOMICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditionally, the cold storage method is used to preserve feces, that is, the feces are collected and stored at a temperature of -20°C or lower, so that the intestinal microorganisms in them are at a low metabolic rate, and the number and composition of the flora are stabilized for a period of time. However, the cold storage method It is still limited by sampling conditions, which requires higher costs for sample transportation, and the quality of storage will be greatly affected by whether it is refrigerated in time

Method used

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  • Excrement preserving fluid and applications thereof
  • Excrement preserving fluid and applications thereof
  • Excrement preserving fluid and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A stool preservation solution (hereinafter referred to as preservation solution 1) includes: ethanol 60% (v / v), sodium citrate 0.74% (w / v), EDTA-disodium 1.5% (w / v), Tris-HCl 10mM , NaCl 0.9% (w / v), sodium lauryl sulfate 0.05% (w / v), and the balance is deionized water. Dissolve the insoluble components in the formula with deionized water, add ethanol, and finally use deionized water to make up the volume, adjust the pH to 8.0, and then sterilize.

Embodiment 2

[0024] A feces preservation solution (hereinafter referred to as preservation solution 2) includes: ethanol 70% (v / v), sodium citrate 1% (w / v), EDTA-disodium 1.86% (w / v), Tris-HCl 10mM , NaCl 0.75% (w / v), sodium lauryl sulfate 0.02% (w / v), and the balance is deionized water. The alcohol-insoluble components in the formula are dissolved with deionized water, then ethanol is added, and finally the volume is adjusted with deionized water, the pH is adjusted to 7.5, and then sterilized.

experiment example

[0034] Taking a healthy volunteer as an example, the mid-stool sample (>10g) is put into the collection box, and 200mg of fresh sample is taken for follow-up testing as the initial test result. Solution 1 group, preservation solution 2 group, contrast solution 1 group, contrast solution 2 group, contrast solution 3 group, Nuohui preservation solution group, add 4mL corresponding preservation solution to 1g of feces sample in each of the above groups, and store at room temperature; -80 In the ℃ group, 1g of feces samples were taken and stored directly at -80°C; a unified test was performed on 7d, 14d, and 30d. For samples without preservative solution, 1g of feces samples were mixed with 4mL of sterile deionized water before testing.

[0035] Test 1. Fecal DNA Extraction

[0036] Using the Tiangen Feces DNA Extraction Kit, the specific steps are as follows:

[0037] 1. Shake and mix the sample, draw 750ul sample into a 2mL centrifuge tube, 5000rpm for 5min;

[0038] 2. Discar...

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Abstract

The invention discloses an excrement preserving fluid and applications thereof. Ethanol is taken as a fixing agent, sodium citrate is taken as an auxiliary fixing agent, EDTA-disodium is taken as an anti-coagulant, Tris-HCl is taken as a buffer solution, NaCl is taken as an ion strength maintaining agent, and the preserving fluid also comprises lauryl sodium sulfate. Through a reasonable formula,the number, composition, and abundance of microbes in an excrement sample are stabilized; the accuracy of sequencing analysis on human intestinal flora is guaranteed; the preserving fluid can preservean excrement sample at a room temperature; compared with a conventional preserving fluid, the preserving effect is better, moreover, provided preserving fluid does not contain any toxic component such as guanidinium thiocyanate, and the operation is safer.

Description

technical field [0001] The invention belongs to the field of microbial detection, and relates to a feces preservation solution and an application thereof. Background technique [0002] Intestinal microorganisms refer to the large number of microorganisms in the intestines of animals. This group of microorganisms lives in the intestines of animals and helps the host to complete various physiological and biochemical functions. Traditional intestinal microbiome research mainly uses methods such as DGGE, FISH, and qPCR. With the development of sequencing technology, intestinal metagenomic sequencing has become a powerful means to study intestinal microbes. Intestinal metagenomic sequencing can analyze the composition, diversity and relative abundance of intestinal flora, and then analyze the relationship between microorganisms and intestinal microenvironment, diseases, drugs, and treatments, and also discover genes with specific functions. Intestinal metagenomic sequencing is d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/04
CPCC12N1/04
Inventor 糜庆丰黄铨飞肖婷婷吴春求夏渝东郭煜王毅王静刘丽菲
Owner CAPITALBIO GENOMICS
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