CN112226369A - Intestinal flora preservation solution and preparation method thereof - Google Patents
Intestinal flora preservation solution and preparation method thereof Download PDFInfo
- Publication number
- CN112226369A CN112226369A CN202011146500.8A CN202011146500A CN112226369A CN 112226369 A CN112226369 A CN 112226369A CN 202011146500 A CN202011146500 A CN 202011146500A CN 112226369 A CN112226369 A CN 112226369A
- Authority
- CN
- China
- Prior art keywords
- preservation solution
- intestinal flora
- intestinal
- sample
- preservative
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/04—Preserving or maintaining viable microorganisms
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Analytical Chemistry (AREA)
- Crystallography & Structural Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Plant Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention provides an intestinal flora preservation solution which is easy for intestinal microorganism collection and long-distance transportation and can maintain the flora abundance and the genetic material stability and a preparation method thereof, and the preparation method comprises the following steps: 0.1-1% (w/v) of preservative, 5-20% (w/v) of fixative, 1.5% (w/v) of antioxidant, 100mM of anticoagulant, 100mM of buffer, 25% (w/v) of ionic strength maintaining agent, and the balance being ddH2And O. The defects that in the prior art, the storage time for collecting the DNA of the excrement sample is short, and the excrement sample needs to be stored and transported at low temperature are effectively overcome, and the method is suitable for industrial large-scale production and application.
Description
Technical Field
The invention is applied to the field of biotechnology, and particularly relates to an intestinal flora preserving fluid and a preparation method thereof.
Background
The intestine is the main organ for digestion and absorption by the human body, and its convoluted structure is figuratively called the second brain of the human body. In the human intestinal tract, a large number of microorganisms including bacteria, fungi and the like are parasitic, wherein 99% of the bacteria account for about 10 trillion bacteria, and the bacteria are called intestinal flora. They are the most important external environment in human body, and various microorganisms are combined according to a certain proportion, are mutually restricted and mutually dependent, and form an ecological balance in quality and quantity. The number of genes of the intestinal flora is about 100 times that of the human body, and therefore is also called "second genome" of the human body. The intestinal flora can not only influence the digestion and absorption of human body to protein, lipid, carbohydrate and other nutrients, but also synthesize various vitamins necessary for the growth and development of human body, regulate the immunity of the organism, resist the risks of infection and autoimmune diseases, closely relate to dozens of diseases such as obesity, type 2 diabetes, colorectal cancer, inflammatory bowel disease and the like, and can control the reaction of the human body to cancer treatment drugs. Under normal conditions, all strains in the intestinal tract are interdependent and mutually restricted with a host, and a dynamic balance is maintained; once the environment inside and outside the body is changed (such as using drugs and the like), the balance state is broken, which causes the imbalance of the intestinal flora and causes various diseases, such as digestive system diseases, endocrine system diseases, mental system diseases, autoimmune diseases and some infectious diseases. It is thus clear that the intestinal flora plays an essential role in maintaining the health of the body.
With the development of biotechnology and genetic testing technology, the interest in the "intestinal flora" of the human second genome is increasing. Due to the wide variety of intestinal microorganisms, it is very important to study the species composition and the original proportion of the intestinal microorganisms and the influence of the intestinal microorganisms on the human body. The 16S rDNA is a gene of prokaryotes coding for 16S rRNA, is about 1500bp in length, and consists of 10 conserved regions and 9 variable regions (V1-V9), wherein the variable regions reflect the difference between species. The method adopts a high-throughput amplicon sequencing technology to detect the specific hypervariable region of the microorganism 16S rRNA gene, and is suitable for identifying the relative abundance of bacteria and archaea in samples such as environmental microorganisms, intestinal flora and the like. The 16S rDNA-seq is obtained by extracting DNA of microbial flora, selecting a specific section (V3-V4) of a variable region for PCR amplification, and helping researchers analyze the genetic composition and function of the microbial flora and the diversity and abundance of the microbial flora in a specific environment by a high-throughput sequencing method, and further analyze the relationship between the microbes and the environment and between the microbes and a host to find genes with specific functions. And detecting the species and abundance of the intestinal microorganisms in the sample by using a 16s technology, guiding the examinee to know the intestinal microecological health condition, evaluating the related disease risk, and customizing a personalized health management scheme. In the medical field, the method is mainly applied to the correlation analysis of intestinal microorganisms and diseases, the microbial difference between the diseases and healthy individuals is revealed, the flora difference after drug or diet interference is researched, or the change of intestinal flora in a certain course of disease development, such as diarrhea, enteritis, colorectal cancer and the like.
The majority of the fecal bacteria are anaerobes, some of which are strictly anaerobes, which easily lose their activity in aerobic environments. In the traditional method, bacteria are separated and cultured by using a solid or liquid culture medium, the period is long, all intestinal flora is difficult to separate and culture due to technical influence, and the research of people on the intestinal flora is limited. At present, the NGS high-throughput sequencing technology is adopted, separation culture is not needed, and the types and the number of the intestinal flora of the human body can be comprehensively detected by extracting and detecting DNA of a fecal sample. Due to the instability of microorganism samples and the complexity of sample preservation, the normal temperature preservation and collection of microorganisms are a common concern. Therefore, the preservation of fecal samples is more critical than the preservation of other bacteria.
At present, the cold storage method is generally adopted clinically for directly storing human excrement, namely, the excrement is frozen at the temperature of minus 20 ℃ or lower immediately after being collected, and the bacterial metabolism rate is reduced in a low-temperature state, so that the composition of the bacteria in the excrement can be maintained for a period of time, and the aim of stabilizing the microbial composition of the excrement is fulfilled. When FMT operation is required, the frozen products are separated to obtain the fecal strain, so that the difficulty and the workload of fecal strain separation are increased, and a larger storage space is also required. In addition, fecal bacteria, as a "drug" in a treatment method, cannot be directly used in clinic, but needs to be frozen and then extracted, and such a treatment process will undoubtedly increase the operation and equipment dependency, and will hinder the development of a fecal bacteria transplantation treatment method. Meanwhile, many medical institutions in China do not have the capability of separating the fecal bacteria. Therefore, although a feces preservation solution (CN104480012A) of patent literature reports that a feces sample can be preserved for one week at a sealed room temperature by using a preservation solution prepared from EDTA salt, sodium citrate, or the like and using water and ethanol as solvents, this method is not suitable for preserving fecal bacteria in fecal bacteria transplantation. At present, Phosphate Buffer Solution (PBS) is often mixed with donor fecal bacteria in the fecal bacteria transplantation process clinically, and due to the fluid characteristics of the mixed solution, the contact time with the intestinal tract wall is short in the process of spraying under an endoscope or clysis clinically, so that the fecal bacteria planting effect is influenced. If the fecal bacteria liquid needs to be stored, glycerol needs to be added additionally, and the excessive steps are easy to increase the chance of exogenous infection.
Therefore, a reagent formula and a preparation method which are easy for collection, storage and transportation of the intestinal microorganisms are provided, and the research on the intestinal microorganisms is facilitated.
Disclosure of Invention
Aiming at the problems that the coprophilous fungi in coprophilous fungi transplantation are difficult to store and easy to infect in the prior art, the invention provides the intestinal flora storage solution which is easy for intestinal microorganism collection and long-distance transportation and can maintain the flora abundance and the genetic material stability and the preparation method thereof.
The technical problem to be solved by the invention is realized by adopting the following technical scheme:
an intestinal flora preservative fluid, which is characterized by comprising:
further, the pH value of the preservation solution is 7.5.
Further, the preservative is selected from alcohols or aldehydes, and preferably, the preservative is hydroxybenzoate or chlorobutanol.
Further, the fixing agent is selected from sucrose or ethanol, the antioxidant is selected from thiourea, the anticoagulant is selected from EDTA-disodium, the buffer is selected from Tris-HCl, and the ionic strength maintaining agent is selected from DMSO.
Preferably, the fixing agent is sucrose. The ethanol can stably preserve the microbial colony structure in the excrement sample for a long time, but the ethanol is inflammable and is not easy to be used for sample transportation, so that the preservation solution needing to be transported selects sucrose as a fixing agent.
DMSO is used as an ionic strength maintaining agent in a certain proportion, and fresh samples stored by DMSO in the proportion can be stored for one day in high-temperature weather at the temperature of not more than 37 ℃ or stored for two weeks at the room temperature of 25 ℃; or storing in a refrigerator at 4 ℃ for one month; or long-term storage at-20 ℃ to keep the DNA intact from degradation. Therefore, the preservative fluid of the invention can be preserved for two weeks at normal temperature without being preserved by freezing. The hydroxybenzoate or chlorobutanol is added as antiseptic to stabilize flora.
Further, an intestinal flora preservation solution comprises:
further, an intestinal flora preservation solution comprises:
further, an intestinal flora preservation solution comprises:
a preparation method of the intestinal flora preservation solution comprises the following steps: weighing the components according to the formula, adding the anticoagulant into ddH2Completely dissolving the O; in ddH2Adding a preservative, a fixing agent, an antioxidant, a buffer solution and an ionic strength maintaining agent into the O, fully dissolving, adding a dissolved anticoagulant, and fully mixing; adjusting pH of the solution to 7.5 and adding ddH2And (4) fixing the volume of O to the required volume to obtain the preservation solution.
Further, the preservative is selected from alcohols or aldehydes, and preferably, the preservative is hydroxybenzoate or chlorobutanol.
Further, the fixing agent is selected from sucrose or ethanol, the antioxidant is selected from thiourea, the anticoagulant is selected from EDTA-disodium, the buffer is selected from Tris-HCl, and the ionic strength maintaining agent is selected from DMSO.
A preservation method of the intestinal flora preservation solution comprises the following steps:
1) collecting at least 1g of a fresh stool sample or an intestinal content sample;
2) soaking the collected sample in a preservation solution according to the proportion of the sample to the preservation solution being 1:3 (w/v);
3) and (3) as soon as possible, transporting the preservation solution soaked with the sample in the step 2) to a laboratory for nucleic acid extraction or preserving at normal temperature/ultralow temperature for later use.
Compared with the prior art, the invention has the beneficial effects that:
the invention can directly put the collected feces or intestinal contents into the preservation solution without freezing and can achieve the effect of stabilizing the microbial composition in the feces or intestinal contents. The preservation solution for the fecal sample or the intestinal tract content sample has high DNA yield of the sample and good sequencing data quality, ensures that the species and the abundance of flora in the sample are consistent with the abundance of flora in a fresh sample, can be transported at normal temperature when stored in the preservation solution, does not need to be frozen, and can be preserved at normal temperature for 2 weeks. The formula of the invention can effectively store DNA in the excrement-like sample for a long time at room temperature, effectively overcomes the defects of short storage time for collecting excrement sample DNA and low-temperature storage and transportation requirements in the prior art, and is suitable for industrial large-scale production and application.
Drawings
FIG. 1 is a statistical chart of 10 samples of OTUs in an experimental example of the preservation solution for intestinal flora and the preparation method of the preservation solution.
Fig. 2 is a bar graph of the relative abundance of species at the level of 10 phyla samples in the experimental example of the preservation solution of intestinal flora and the preparation method of the preservation solution.
Fig. 3 shows diversity analysis of 10 samples in experimental examples of the preservation solution of intestinal flora and the preparation method of the preservation solution.
Detailed Description
The technical solution of the present invention is further described in detail below with reference to the accompanying drawings and specific embodiments. It is to be understood that the following examples are only illustrative and explanatory of the present invention and should not be construed as limiting the scope of the present invention. All the technologies realized based on the above-mentioned contents of the present invention are covered in the protection scope of the present invention.
In addition, unless otherwise specifically indicated, various starting materials, reagents, instruments and equipment used in the present invention may be commercially available or prepared by existing methods.
Example 1:
an intestinal flora preservative fluid comprising:
preparing an intestinal flora preservation solution:
weighing the components according to the formula, adding EDTA-disodium into ddH2Completely dissolving the O; in ddH2Adding hydroxyl benzoate, sucrose, thiourea, Tris-HCl and DMSO into the O, fully dissolving, adding the dissolved anticoagulant, and fully mixing; adjusting pH of the solution to 7.5 and adding ddH2And (4) fixing the volume of O to the required volume to obtain the preservation solution.
Example 2:
an intestinal flora preservative fluid comprising:
preparing an intestinal flora preservation solution:
weighing the components according to the formula, adding EDTA-disodium into ddH2Completely dissolving the O; in ddH2Adding hydroxyl benzoate, sucrose, thiourea, Tris-HCl and DMSO into the O, fully dissolving, adding the dissolved anticoagulant, and fully mixing; adjusting pH of the solution to 7.5 and adding ddH2And (4) fixing the volume of O to the required volume to obtain the preservation solution.
Example 3:
an intestinal flora preservative fluid comprising:
preparing an intestinal flora preservation solution:
weighing the components according to the formula, adding EDTA-disodium into ddH2Completely dissolving the O; in ddH2Adding chlorobutanol, absolute ethyl alcohol, thiourea, Tris-HCl and DMSO into the O, fully dissolving, adding the dissolved anticoagulant, and fully mixing; adjusting pH of the solution to 7.5 and adding ddH2And (4) fixing the volume of O to the required volume to obtain the preservation solution.
Examples of the experiments
1. Preparation of preservative fluid
Preparing a preservation solution according to the scheme of the embodiment 1 of the invention for later use, and naming the preservation solution as CN-2; preparing a preservation solution according to the scheme of patent CN 104480012A:
weighing the components according to the formula, dissolving the components in sterile deionized water, sterilizing at high pressure, and storing at room temperature for later use, wherein the name of the component is CN-1.
2. Sample collection and processing
Stool samples from the same individual were collected and divided into 10 equal portions, each greater than 1g, and numbered ZS-1, ZS-2, ZS-3, ZS-4, ZS-10, ZS-11, ZS-12, ZS-14, ZS-15, and ZS-16, respectively. The sample treatment was as follows:
| sample numbering | Sample processing method |
| ZS-1 | Directly extracting intestinal microorganism DNA within 10 minutes without adding any preservation solution for later use |
| ZS-2 | Storing at room temperature for 24 hr without adding any preservative solution, extracting intestinal microorganism DNA, and keeping |
| ZS-10 | Storing at room temperature for 3 × 24 hr without adding any preserving solution, extracting intestinal microorganism DNA, and keeping |
| ZS-14 | Storing at room temperature for 7 × 24 hr without adding any preserving solution, extracting intestinal microorganism DNA, and keeping |
| ZS-3 | Adding CN-1 preserving fluid, preserving at room temperature for 24 hr, extracting intestinal microorganism DNA, and keeping |
| ZS-11 | Adding CN-1 preserving fluid, preserving at room temperature for 3 × 24 hr, extracting intestinal microorganism DNA, and keeping |
| ZS-15 | Adding CN-1 preserving fluid, preserving at room temperature for 7 × 24 hr, extracting intestinal microorganism DNA, and keeping |
| ZS-4 | Adding CN-2 preserving fluid, preserving at room temperature for 24 hr, extracting intestinal microorganism DNA, and keeping |
| ZS-12 | Adding CN-2 preserving solution, preserving at room temperature for 3 × 24 hr, and extracting intestinal microorganismsDNA, ready for use |
| ZS-16 | Adding CN-2 preservation solution, preserving at room temperature for 7 × 24 hr, extracting intestinal microorganism DNA, and keeping |
The preservation method for adding the preservation solution into the samples ZS-3, ZS-11, ZS-15, ZS-4, ZS-12 and ZS-16 is as follows: the proportion of the amount of each sample to the preservation solution is 1:3(w/v), each sample is soaked in the preservation solution according to the proportion, and a disposable 10ml sterilization tube is adopted for room temperature preservation.
3. Sample DNA extraction
In the step 2, the fecal DNA extraction adopts a fecal DNA extraction kit (product number: D3141) produced by Meiji organisms, and the operation steps are carried out according to the instruction. The method comprises the following specific steps:
transferring 150-200 mg of the fecal sample into a 2ml centrifuge tube. 1.2ml Buffer SSL was immediately added to the sample and the sample was thoroughly broken up by vortexing for a maximum of 1 minute. Water bath at 70 ℃ for 10 min. Vortex for 15 seconds. Centrifuging at room temperature at 14000 Xg for 10 min. Transfer 250. mu.l of supernatant to a new 1.5ml centrifuge tube. Add 20. mu.l of Proteinase K and 250. mu.l of Buffer AL supernatant. Mix by inversion 10 times. Water bath at 70 ℃ for 10 min. Add 250. mu.l of absolute ethanol to the sample and mix 10 times by inversion. HiPure DNA Mini Column I was loaded into a 2ml collection tube. Transfer the mixture to the column. Centrifuging at 10000 Xg for 30-60 seconds. The effluent is decanted and the column is returned to the collection tube. Mu.l of Buffer GW1 (diluted with absolute ethanol) was added to the column. Centrifuging at 10000 Xg for 30-60 seconds. The filtrate was decanted and the column was returned to the collection tube. 650. mu.l of Buffer GW2 (diluted with ethanol) was added to the column. Centrifuging at 10000 Xg for 30-60 seconds. Buffer GW2 was diluted with absolute ethanol. The filtrate was decanted and the column was returned to the collection tube. 13000 Xg centrifuge for 2 minutes to spin dry the column. The column was placed in a 1.5ml centrifuge tube. 50-200. mu.l of Buffer AE preheated to 70 ℃ was added to the center of the membrane of the column and left at room temperature for 2 minutes. 13000 Xg for 1 minute. The DNA binding column was discarded and the DNA was stored at-20 ℃.
4. 16S rRNA sequencing
The 10 samples were subjected to 16S rRNA sequencing and the differences and changes in the bacterial population structure of the 10 samples were compared by analysis. In this example, the V3-V4 region of the microorganism 16SrRNA was examined. Sequencing was performed using a Miseq high throughput sequencer produced by Illumina.
5. Structural analysis of flora
And (3) obtaining floras corresponding to 10 samples through 16SrRNA sequencing data and bioinformatics analysis, and calculating species diversity indexes for comparison. Referring to table 1, the present inventors counted the sequencing result of sample No. 10 and evaluated the sequencing quality to meet the analysis requirements. The present invention analyzed OTUs statistics and bacterial relative abundance composition (see fig. 1, fig. 2) and microbiota diversity composition (see fig. 3) at the phylum level for 10 samples.
Referring to Table 1 and accompanying figures 1-3, based on the analysis of bacterial abundance, flora relationships, and genus composition, the formula preservation solution of the present invention showed little change in bacterial abundance over time (ZS-4, ZS-12, ZS-16), while the preservation solution CN-1 showed a greater change in bacterial abundance at day 3 (ZS-3, ZS-11, ZS-15), while the sample without any preservation solution showed a greater change in bacterial abundance; it can be seen that the preservation solution of the invention has better effect of maintaining the relative abundance of bacteria compared with the preservation solution CN-1 without adding any preservation solution. The flora diversity of the formula preservation solution of the invention is changed from 6.36 to 6.19 on day 7, and the flora diversity of the preservation solution CN-1 is changed from 6.25 to 5.96 on day 7, although the difference between the flora diversity of the preservation solution and the flora diversity of the preservation solution is small after day 1, the flora diversity of the frozen stock solution is greatly changed along with the time, and the formula preservation solution of the invention is better than the traditional frozen stock solution in the aspect of flora diversity.
The statistical result shows that the preservation liquid CN-2 is adopted for normal-temperature transportation and preservation, and the high-efficiency stabilization period can reach 7 days. Compared with the traditional frozen stock solution (CN-1) treatment, the effect of the preservation solution formula of the invention is obviously superior and is obviously superior to the preservation scheme without any treatment in the experimental examples.
Table 110 sample 16SrRNA sequencing quality statistics table
The above description is only a preferred embodiment of the present invention, and the protection scope of the present invention is not limited to the above-described embodiments. It will be understood by those skilled in the art that various changes, substitutions of equivalents, and alterations can be made without departing from the spirit and scope of the invention.
Claims (9)
1. An intestinal flora preservative fluid, comprising:
2. the intestinal flora preservative fluid according to claim 1, wherein: the pH value of the preservation solution is 7.5.
3. The intestinal flora preservative fluid according to claim 1, wherein: the preservative is selected from alcohols or aldehydes, preferably hydroxybenzoate or chlorobutanol.
4. The intestinal flora preservative fluid according to claim 1, wherein: the fixing agent is selected from sucrose or ethanol, the antioxidant is selected from thiourea, the anticoagulant is selected from EDTA-disodium, the buffer is selected from Tris-HCl, and the ionic strength maintaining agent is selected from DMSO.
5. The intestinal flora preservative fluid according to claim 1, comprising:
6. the intestinal flora preservative fluid according to claim 5, comprising:
7. the intestinal flora preservative fluid according to claim 6, comprising:
8. a method for producing a preservation solution for intestinal flora according to any one of claims 1 to 7, wherein the preservation solution comprises: weighing the components according to the formula, adding the anticoagulant into ddH2Completely dissolving the O; in ddH2Adding a preservative, a fixing agent, an antioxidant, a buffer solution and an ionic strength maintaining agent into the O, fully dissolving, adding a dissolved anticoagulant, and fully mixing; adjusting pH of the solution to 7.5 and adding ddH2And (4) fixing the volume of O to the required volume to obtain the preservation solution.
9. A method for preserving an intestinal flora preservative fluid according to claim 8, wherein the method comprises the steps of:
1) collecting at least 1g of a fresh stool sample or an intestinal content sample;
2) soaking the collected sample in a preservation solution according to the proportion of the sample to the preservation solution being 1:3 (w/v);
3) and (3) as soon as possible, transporting the preservation solution soaked with the sample in the step 2) to a laboratory for nucleic acid extraction or preserving at normal temperature/ultralow temperature for later use.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011146500.8A CN112226369A (en) | 2020-10-23 | 2020-10-23 | Intestinal flora preservation solution and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011146500.8A CN112226369A (en) | 2020-10-23 | 2020-10-23 | Intestinal flora preservation solution and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112226369A true CN112226369A (en) | 2021-01-15 |
Family
ID=74109384
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011146500.8A Pending CN112226369A (en) | 2020-10-23 | 2020-10-23 | Intestinal flora preservation solution and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112226369A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114469852A (en) * | 2022-02-18 | 2022-05-13 | 厦门承葛生物科技有限公司 | Preparation method of intestinal flora gel for flora transplantation |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105543095A (en) * | 2016-03-04 | 2016-05-04 | 微基生物科技（上海）有限公司 | Preservation reagent for microorganism specimens containing rich organic matters, kit containing preservation reagent and application of preservation reagent |
| CN107312717A (en) * | 2017-08-08 | 2017-11-03 | 苏州阿迪康生物科技有限公司 | A kind of store method of excrement class sample, preservation solution, preparation method and application |
| CN107530280A (en) * | 2015-04-24 | 2018-01-02 | 玛阿特制药公司 | The preparation method of fecal microorganism group's sample |
| CN107988076A (en) * | 2017-12-19 | 2018-05-04 | 东莞博奥木华基因科技有限公司 | A kind of excrement preserves liquid and its application |
-
2020
- 2020-10-23 CN CN202011146500.8A patent/CN112226369A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107530280A (en) * | 2015-04-24 | 2018-01-02 | 玛阿特制药公司 | The preparation method of fecal microorganism group's sample |
| CN105543095A (en) * | 2016-03-04 | 2016-05-04 | 微基生物科技（上海）有限公司 | Preservation reagent for microorganism specimens containing rich organic matters, kit containing preservation reagent and application of preservation reagent |
| CN107312717A (en) * | 2017-08-08 | 2017-11-03 | 苏州阿迪康生物科技有限公司 | A kind of store method of excrement class sample, preservation solution, preparation method and application |
| CN107988076A (en) * | 2017-12-19 | 2018-05-04 | 东莞博奥木华基因科技有限公司 | A kind of excrement preserves liquid and its application |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114469852A (en) * | 2022-02-18 | 2022-05-13 | 厦门承葛生物科技有限公司 | Preparation method of intestinal flora gel for flora transplantation |
| CN114469852B (en) * | 2022-02-18 | 2024-04-26 | 厦门承葛生物科技有限公司 | Preparation method of intestinal flora gel for flora transplantation |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP6858747B2 (en) | How to prepare a fecal microbiota sample | |
| Callegari et al. | Compartmentalization of bacterial and fungal microbiomes in the gut of adult honeybees | |
| AU2019253714A1 (en) | Microbial consortia | |
| Gorrens et al. | Isolation and identification of dominant bacteria from black soldier fly larvae (Hermetia illucens) envisaging practical applications | |
| Weir | The ecology of Zymomonas: a review | |
| CN109589275A (en) | Ectoin is maintaining the purposes in skin microecological balance | |
| CN114369537A (en) | Excrement storage liquid and preparation method and application thereof | |
| Yang et al. | The beneficial effects of polysaccharide obtained from persimmon (Diospyros kaki L.) on the proliferation of Lactobacillus and gut microbiota | |
| Rezaei et al. | Isolation of lactic acid probiotic strains from Iranian camel milk: technological and antioxidant properties | |
| CN112226369A (en) | Intestinal flora preservation solution and preparation method thereof | |
| CN114574393A (en) | Lactobacillus delbrueckii SEUNEU-110 and application thereof in skin | |
| Zheng et al. | Lactic acid bacteria are prevalent in the infrabuccal pockets and crops of ants that prefer aphid honeydew | |
| Kozhakhmetov et al. | Gut modulation of dysbiosis induced by dextran sulfate sodium | |
| Kirschner et al. | Characterization of fungal and bacterial components in gut/fecal microbiome | |
| JP2022130286A (en) | Compositions for improving bacterial flora, methods for improving bacterial flora using same, and applications thereof | |
| JP2024501818A (en) | How to grow complex communities of microorganisms | |
| CN113122473A (en) | Lactobacillus BB1 for enhancing memory capacity, fermented food thereof and application thereof | |
| Domínguez et al. | Metagenomic analysis of the intestinal microbiome in goats on cactus and Salicornia-based diets | |
| EP3974536A1 (en) | Bacterium provision system | |
| Diyaolu et al. | Phenotypic and molecular characterization of different isolates of Lactobacillus plantarum from four Nigerian fermented foods for use as probiotics in aquaculture | |
| Ferreira de Vilcinskas | Identification of Lactobacillus species in honey using 16S rRNA gene sequencing | |
| CN110616163B (en) | Enterococcus faecalis capable of reducing concentration of trimethylamine and preparation method and application thereof | |
| CN112300962B (en) | Lactobacillus plantarum with specific molecular target and antioxidation function and application thereof | |
| Chang et al. | Isolation, Identification and Biological Characteristics of Lactobacillus reuteri from Tibetan Pigs | |
| Vernero et al. | 9 Challenge, Future Research, and Influence in Food Science and Product Development |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210115 |
|
| RJ01 | Rejection of invention patent application after publication |