CN114480129A - Human excrement preserving fluid and using method thereof - Google Patents
Human excrement preserving fluid and using method thereof Download PDFInfo
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- CN114480129A CN114480129A CN202210034812.2A CN202210034812A CN114480129A CN 114480129 A CN114480129 A CN 114480129A CN 202210034812 A CN202210034812 A CN 202210034812A CN 114480129 A CN114480129 A CN 114480129A
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- excrement
- human
- preservation solution
- human excrement
- solution
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- 238000000034 method Methods 0.000 title claims abstract description 7
- 239000012530 fluid Substances 0.000 title description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims abstract description 9
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims abstract description 6
- FSYKKLYZXJSNPZ-UHFFFAOYSA-N sarcosine Chemical compound C[NH2+]CC([O-])=O FSYKKLYZXJSNPZ-UHFFFAOYSA-N 0.000 claims abstract description 6
- 239000011780 sodium chloride Substances 0.000 claims abstract description 5
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims abstract description 4
- 229940098773 bovine serum albumin Drugs 0.000 claims abstract description 4
- 239000004471 Glycine Substances 0.000 claims abstract description 3
- 108010077895 Sarcosine Proteins 0.000 claims abstract description 3
- 239000002518 antifoaming agent Substances 0.000 claims abstract description 3
- 229940043230 sarcosine Drugs 0.000 claims abstract description 3
- 239000003761 preservation solution Substances 0.000 claims description 11
- 239000000243 solution Substances 0.000 claims description 8
- 239000008367 deionised water Substances 0.000 claims description 2
- 229910021641 deionized water Inorganic materials 0.000 claims description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 2
- 239000007788 liquid Substances 0.000 abstract description 5
- 244000005700 microbiome Species 0.000 description 8
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 150000007523 nucleic acids Chemical class 0.000 description 3
- 102000039446 nucleic acids Human genes 0.000 description 3
- 108020004707 nucleic acids Proteins 0.000 description 3
- 238000007400 DNA extraction Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000002550 fecal effect Effects 0.000 description 2
- 210000003608 fece Anatomy 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 230000002335 preservative effect Effects 0.000 description 2
- 238000000018 DNA microarray Methods 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 1
- 229930182566 Gentamicin Natural products 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 239000002981 blocking agent Substances 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 229960002518 gentamicin Drugs 0.000 description 1
- YQOKLYTXVFAUCW-UHFFFAOYSA-N guanidine;isothiocyanic acid Chemical compound N=C=S.NC(N)=N YQOKLYTXVFAUCW-UHFFFAOYSA-N 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 239000000565 sealant Substances 0.000 description 1
- 210000001082 somatic cell Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
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
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to the technical field of biological application, in particular to human excrement storage liquid and a using method thereof, wherein the human excrement storage liquid comprises the following components: 6.5-8g/L glycine, 8-9.5g/L sodium chloride, 5-6g/L bovine serum albumin, 200-220g/L sarcosine, 0.01-0.1% Krovin500, 1-5mL10M NaOH solution, 300 muL/L antifoaming agent SE-15. Provides the excrement storage liquid which is safe and convenient and can be well stored and transported at normal temperature.
Description
Technical Field
The invention relates to the technical field of biological application, in particular to human excrement storage liquid and a using method thereof.
Background
The number of human microorganisms is huge, the number is 10 times higher than the number of human somatic cells, the microorganisms are considered as an 'organ' of human body by some researchers, the 'organ' is concerned with the health of human body, the detection of microorganisms is more and more important, most of the microorganisms exist in intestinal tract, and therefore, the DNA of the microorganisms needs to be extracted from excrement so as to carry out related detection. The existing problem is that once the excrement is discharged, the living environment of microorganisms is changed greatly, so the excrement needs to be stored in time, and the microorganisms in the excrement can be well stored at the temperature of minus 20 ℃ for 1 week, but in many cases, the excrement sample is not collected in a laboratory and cannot be stored in the environment of minus 20 ℃ in time, so a storage solution capable of storing the excrement microorganisms at normal temperature and a corresponding using method are needed.
Chinese patent application publication No. CN 106769289 a discloses a composition comprising "buffer: 20-50mM Tris buffer (pH 7.4), blocking agent: 5-15% bovine serum albumin, 0.10-0.20% Tween-20, 120-: 10-20mM EDTA, preservative: 0.090-0.15% sodium azide and 10-20 mug/mL gentamicin feces preservation solution, although the common sealant ethanol is replaced for convenient transportation, the use of the highly toxic sodium azide as the preservative still has great risk for medical care personnel.
Disclosure of Invention
The invention aims to solve the defects of the prior art and provide the excrement storage liquid which is safe and convenient and can be well stored and transported at normal temperature.
In order to achieve the above object, a human feces preservation solution is designed, comprising: 6.5-8g/L glycine, 8-9.5g/L sodium chloride, 5-6g/L bovine serum albumin, 200-220g/L sarcosine, 0.01-0.1% Krovin500, 1-5mL of 10M NaOH solution, and 300 muL/L defoaming agent SE-15.
Preferably, all components of the preservation solution are dissolved with sterile deionized water.
Preferably, the pH of the preservation solution is 8.0-8.5.
Also relates to a using method of the preservation solution, when in use, the mass volume ratio of the fresh excrement to the preservation solution is 1: 3.
compared with the prior art, the invention has the advantages that: the DNA-free DNA chip is free of ethanol, convenient to transport, free of guanidine isothiocyanate, sodium azide and other toxic substances, safer, and capable of being extracted in large quantity, and DNA is not damaged after being stored for one week at normal temperature.
Detailed Description
The present invention is further described below in conjunction with specific embodiments, the structure and principles of which will be apparent to those skilled in the art. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1: (this formulation)
Taking fresh excrement and preserving fluid according to the mass-volume ratio of 1: 3, mixing, storing at normal temperature for one week, and extracting DNA by using a fecal genome DNA extraction kit (centrifugal column type) of Tiangen.
Comparative example 1: (physiological saline control group)
Taking fresh excrement and 0.9% sodium chloride solution according to the mass-volume ratio of 1: 3, mixing, storing at normal temperature for one week, and extracting DNA by using a fecal genome DNA extraction kit (centrifugal column type) of Tiangen.
Different testing of the nucleic acids extracted in example 1 and comparative example 1 above:
the concentrations of the nucleic acids of example 1 and comparative example 1 were determined using an ultraviolet spectrophotometer, and the experimental results are shown below:
TABLE 1 results of concentration and purity measurement of nucleic acids extracted in different examples
Comparing example 1 with comparative example 1, it can be seen that the concentration of DNA extracted after the sample is stored for one week in example 1 is 213.1ng/mL, while the concentration of DNA extracted after the sample is stored for one week in comparative example 1 is 7.8ng/mL, which is very different from that in example 1, and it can be seen from the difference between the two concentrations that the sample stored by the storage solution can still extract DNA with high concentration after one week, while the DNA is seriously damaged after one week in the sample stored by the 0.9% sodium chloride solution, and the rest is very small.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (4)
1. A human stool preservation solution, comprising: 6.5-8g/L glycine, 8-9.5g/L sodium chloride, 5-6g/L bovine serum albumin, 200-220g/L sarcosine, 0.01-0.1% Krovin500, 1-5mL10M NaOH solution, 300 muL/L antifoaming agent SE-15.
2. The human stool preservation solution according to claim 1, wherein all components of the preservation solution are dissolved by sterile deionized water.
3. The human stool preservation solution according to claim 1 or 2, wherein the pH of the preservation solution is 8.0-8.5.
4. The use method of the human excrement storage solution according to claim 1, wherein the mass-to-volume ratio of the fresh excrement to the storage solution is 1: 3.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210034812.2A CN114480129A (en) | 2022-01-13 | 2022-01-13 | Human excrement preserving fluid and using method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210034812.2A CN114480129A (en) | 2022-01-13 | 2022-01-13 | Human excrement preserving fluid and using method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114480129A true CN114480129A (en) | 2022-05-13 |
Family
ID=81512227
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210034812.2A Pending CN114480129A (en) | 2022-01-13 | 2022-01-13 | Human excrement preserving fluid and using method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114480129A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1205628A (en) * | 1995-10-25 | 1999-01-20 | 曼海姆泊灵格股份公司 | Method and prepns. for stabilizing biological materials by drying methods without freezing |
| CN102575286A (en) * | 2009-04-20 | 2012-07-11 | 长角牛疫苗和诊断有限责任公司 | Biological specimen collection/transport compositions and methods |
| RU2619898C1 (en) * | 2016-03-24 | 2017-05-19 | Общество с ограниченной ответственностью "Биологическая среда" | Method for biological materials preservation |
| CN106769289A (en) * | 2016-12-02 | 2017-05-31 | 刘鹏飞 | A kind of collocation method of fecal specimens storage liquid and application |
| CN113637723A (en) * | 2021-07-09 | 2021-11-12 | 江苏康为世纪生物科技股份有限公司 | Fecal sample nucleic acid preservation solution and preparation method thereof |
-
2022
- 2022-01-13 CN CN202210034812.2A patent/CN114480129A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1205628A (en) * | 1995-10-25 | 1999-01-20 | 曼海姆泊灵格股份公司 | Method and prepns. for stabilizing biological materials by drying methods without freezing |
| CN102575286A (en) * | 2009-04-20 | 2012-07-11 | 长角牛疫苗和诊断有限责任公司 | Biological specimen collection/transport compositions and methods |
| RU2619898C1 (en) * | 2016-03-24 | 2017-05-19 | Общество с ограниченной ответственностью "Биологическая среда" | Method for biological materials preservation |
| CN106769289A (en) * | 2016-12-02 | 2017-05-31 | 刘鹏飞 | A kind of collocation method of fecal specimens storage liquid and application |
| CN113637723A (en) * | 2021-07-09 | 2021-11-12 | 江苏康为世纪生物科技股份有限公司 | Fecal sample nucleic acid preservation solution and preparation method thereof |
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| 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: 20220513 |
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| RJ01 | Rejection of invention patent application after publication |