CN109122667A - Save liquid and its preparation method and application - Google Patents
Save liquid and its preparation method and application Download PDFInfo
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- CN109122667A CN109122667A CN201811137087.1A CN201811137087A CN109122667A CN 109122667 A CN109122667 A CN 109122667A CN 201811137087 A CN201811137087 A CN 201811137087A CN 109122667 A CN109122667 A CN 109122667A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0226—Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
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Abstract
The present invention provides a kind of excrement to save liquid and its preparation method and application, Preservation methods of faeces, belongs to biologic product technology field.The present invention provides a kind of excrement to save liquid, and it includes following ingredient that the excrement, which saves liquid: chelating agent 10-500mmol/L, bacteriostatic agent 100-600mL/L, ionic strength maintain agent 1-10g/L and denaturant 1-50g/L.The excrement, which saves, can be reserved for fecal sample under liquid room temperature, and can effectively inhibit DNA degradation and cell damage in fecal sample, maintains cellular morphology, inhibits degrading enzyme and microbial activity effect good, the Sample preservation time is long.
Description
Technical field
The invention belongs to biologic product technology fields, and in particular to a kind of excrement save liquid and its preparation method and application,
Preservation methods of faeces.
Background technique
In recent years, the detection of cast-off cells is usually used in tying due to noninvasive painless and specific high feature in human faecal mass
The primary dcreening operation of the diseases such as the carcinoma of the rectum is generally investigated, more compared with the methods of the occult blood test of chemical method or immunization, full proctoscopy
It can achieve the purpose that " early discovery, early diagnosis, early treatment ".
But excrement ingredient is sufficiently complex, contains a large amount of digestion remnants and bacteriogenic various nucleolysis enzyme such as albumen
Enzyme, DNA enzymatic (DNases) and RNA enzyme (RNases), various PCR inhibitors (such as cholate, bilirubin, humus and pigment), make
Cell and nucleic acid in fecal sample is obtained to be highly prone to destroy and degrade.Currently used store method is by acquiring excrement sample
It is frozen at -20 DEG C or in lower temperature immediately after this, degradation or other biochemical reactions to avoid cast-off cells.But
Fecal sample acquisition is not many times to carry out in the lab, that is to say, that the fecal sample of acquisition more or less all needs
To place a period of time at normal temperature can just be put into -20 DEG C of preservations in refrigerator；This process be easy to cause originally just minimal amount of
Cast-off cells further loss, to influence subsequent detection.
Current fecal sample store method, the storable time is short, and sample utilizes qualification rate low once again, and preservation condition is severe
It carves, it is at high cost.Thus, current fecal sample liquid and store method still have much room for improvement.It is therefore desired to provide a kind of energy
Enough reagents for effectively keeping fecal sample for a long time at normal temperature, the reagent can effectively inhibit in fecal sample DNA degradation and thin
Born of the same parents are impaired, inhibit degrading enzyme and microbial activity, and do not influence excrement detection, are able to solve at least one of above problem.
In consideration of it, the present invention is specifically proposed.
Summary of the invention
The purpose of the present invention is to provide a kind of excrement to save liquid；The excrement saves liquid and can save for a long time at normal temperature
Fecal sample, and can effectively inhibit DNA degradation and cell damage in fecal sample maintains cellular morphology, inhibits degrading enzyme and micro-
Bioactivity effect is good.
The second object of the present invention is to provide the preparation method that above-mentioned excrement saves liquid.
The third object of the present invention is that providing above-mentioned excrement saves application of the liquid in excrement preservation.
The fourth object of the present invention is to provide a kind of Preservation methods of faeces；This method is to save liquid using above-mentioned excrement to protect
It deposits.
According to an aspect of the invention, there is provided a kind of excrement saves liquid, it includes following ingredient that the excrement, which saves liquid:
Chelating agent 10-500mmol/L, bacteriostatic agent 100-600mL/L, ionic strength maintain agent 1-10g/L and denaturant 1-
50g/L。
As further preferred technical solution, it includes following ingredient that the excrement, which saves liquid:
Chelating agent 20-300mmol/L, bacteriostatic agent 200-500mL/L, ionic strength maintain agent 5-10g/L and denaturant 5-
30g/L。
As further preferred technical solution, it includes following ingredient that the excrement, which saves liquid:
Chelating agent 30-100mmol/L, bacteriostatic agent 200-300mL/L, ionic strength maintain agent 8-10g/L and denaturant 5-
20g/L。
As further preferred technical solution, the pH value that the excrement saves liquid is 6.0-11.0, preferably 7.0-
11.0, further preferably 7.0-8.0；
Preferably, the pH that the excrement saves liquid is mediated by the way that buffer is added；
Preferably, the buffer includes Tris-HCl, phosphate buffer, acetate buffer or Na2HPO4Lemon
Sour sodium buffer.
As further preferred technical solution, the chelating agent includes organic sequestering agent；
Preferably, the organic sequestering agent includes edetate, 1,2-cyclohexanediaminetetraacetic acid salt, diethyl three
The double aminoethyl ether tetraacethyls of triamine pentaacetic acid salt, trientine salt, ethylene glycol, diethyldithiocar bamic acid, nitrilo- three
At least one of acetic acid and ethylenediamine disuccinic acid trisodium；
Preferably, the organic sequestering agent include in edetate and 1,2-cyclohexanediaminetetraacetic acid salt at least
It is a kind of.
As further preferred technical solution, the bacteriostatic agent includes substituted or unsubstituted C1-C4Alcohol, wherein described
Substituent group includes fluorine, chlorine or bromine；
Preferably, the bacteriostatic agent includes at least one of methanol, ethyl alcohol, ethylene glycol, isopropanol and methaform, preferably
For at least one of ethyl alcohol and methaform；
Preferably, it includes sodium chloride that the ionic strength, which maintains agent,.
As further preferred technical solution, the denaturant includes anionic surfactant and amphoteric ion surface
At least one of activating agent；
Preferably, the anionic surfactant include sulfate type anionic surfactant and sulfonate type yin from
At least one of sub- surfactant；
Preferably, the sulfate type anionic surfactant includes urea, lauryl sodium sulfate, dodecyl sulphur
Sour ammonium, lithium dodecyl sulfate, sodium decyl sulfate, sodium octyl sulfate, ethoxylated dodecyl alcohol sodium sulfovinate, sulphation castor-oil plant
Oil, lauryl sulfate diethanolamine, dodecyltriethanolamine sulfate, lauryl potassium sulfate, triethanolamine lauryl sulfate, the moon
At least one of osmanthus base diethanolamine and lauryl magnesium sulfate, preferably in urea and lauryl sodium sulfate at least
It is a kind of；
Preferably, the sulfonate type anionic surfactant includes sulfosuccinic acid-diisobutylester-S-sodium salt, sulfosuccinic
Acid -1,4- diamyl ester sodium, sulfo-succinic acid dihexyl sodium, sulfosuccinate dicyclohexyl maleate sodium, hendecene base single ethanol amide amber
At least one of amber acid esters sodium sulfonate, sulfosuccinic acid lauryl disodium and Sodium docusate, preferably sulfo group
Dioctyl succinate sodium；
Preferably, the zwitterionic surfactant includes at least one of chlorination guanidine and guanidine thiocyanate, preferably
Guanidine thiocyanate.
According to the second aspect of the invention, the preparation method that the excrement saves liquid is provided, including by the original
Material is configured to solution, obtains excrement and saves liquid；
Preferably, it after raw material being configured to solution, is filtered using filter membrane device, obtains excrement and save liquid.
According to the third aspect of the present invention, it provides the excrement and saves application of the liquid in excrement preservation.
According to the fourth aspect of the present invention, a kind of Preservation methods of faeces is provided, the Preservation methods of faeces is to use
The excrement saves liquid and saves.
The present invention provides a kind of excrement save liquid, including chelating agent 10-500mmol/L, bacteriostatic agent 100-600mL/L,
Ionic strength maintains agent 1-10g/L and denaturant 1-50g/L.The excrement, which saves, can be reserved for fecal sample under liquid room temperature, and energy
Effectively inhibit DNA degradation and cell damage in fecal sample, maintain cellular morphology, inhibits degrading enzyme and microbial activity effect
Good, the Sample preservation time is long.
Detailed description of the invention
It, below will be to specific in order to illustrate more clearly of the specific embodiment of the invention or technical solution in the prior art
Embodiment or attached drawing needed to be used in the description of the prior art be briefly described, it should be apparent that, it is described below
Attached drawing is some embodiments of the present invention, for those of ordinary skill in the art, before not making the creative labor
It puts, is also possible to obtain other drawings based on these drawings.
Fig. 1 is the agarose gel electrophoresis figure that 1 excrement of the embodiment of the present invention saves that liquid 7 days fecal samples of preservation obtain；
Fig. 2 is the agarose gel electrophoresis figure that negative control group (physiological saline) saves that 7 days fecal samples obtain.
Specific embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person carries out according to conventional conditions or manufacturer's recommended conditions.Reagents or instruments used without specified manufacturer is
The conventional products that can be obtained by commercially available purchase.
According to an aspect of the invention, there is provided a kind of excrement saves liquid, it includes following ingredient that the excrement, which saves liquid:
Chelating agent 10-500mmol/L, bacteriostatic agent 100-600mL/L, ionic strength maintain agent 1-10g/L and denaturant 1-
50g/L。
The excrement, which saves, can be reserved for fecal sample under liquid room temperature, and can effectively inhibit in fecal sample DNA degradation and thin
Born of the same parents are impaired, maintain cellular morphology, inhibit degrading enzyme and microbial activity effect good, the Sample preservation time is long.
It should be noted that the present invention maintains the source of agent and denaturant not have chelating agent, bacteriostatic agent, ionic strength
Special limitation, using each raw material well-known to those skilled in the art；Its commercial goods can be such as used, can also be adopted
It is voluntarily prepared with preparation method well known to those skilled in the art.
Wherein, chelating agent is capable of inhibiting cell impaired, maintains cellular morphology, inhibits the activity of enzyme；Chelating agent typical case but non-limit
The concentration of property processed be 10mmol/L, 20mmol/L, 30mmol/L, 40mmol/L, 50mmol/L, 60mmol/L, 70mmol/L,
80mmol/L、90mmol/L、100mmol/L、120mmol/L、130mmol/L、150mmol/L、180mmol/L、200mmol/
L、220mmol/L、240mmol/L、260mmol/L、270mmol/L、290mmol/L、300mmol/L、320mmol/L、
340mmol/L、360mmol/L、370mmol/L、380mmol/L、400mmol/L、410mmol/L、420mmol/L、
440mmol/L, 460mmol/L, 480mmol/L or 500mmol/L.
Wherein, bacteriostatic agent can prevent the abundance of microorganism from changing, and inhibit the activity of microorganism；Bacteriostatic agent it is typical but
Unrestricted concentration be 100mL/L, 120mL/L, 130mL/L, 150mL/L, 180mL/L, 200mL/L, 220mL/L,
240mL/L、260mL/L、270mL/L、290mL/L、300mL/L、320mL/L、340mL/L、360mL/L、370mL/L、
380mL/L、400mL/L、410mL/L、420mL/L、440mL/L、460mL/L、480mL/L、500mL/L、510mL/L、
520mL/L, 540mL/L, 560mL/L, 580mL/L or 600mL/L.
Ionic strength maintenance agent can maintain the stabilization of total ionic strength；Ionic strength maintains agent typical but non-limiting dense
Degree is 1g/L, 2g/L, 3g/L, 4g/L, 5g/L, 5.5g/L, 6g/L, 7g/L, 8g/L, 9g/L, 9.5g/L or 10g/L.
Denaturant can by micro- fecal specimens mucoprotein and the organic substances such as globulin carry out denaturation treatment；Denaturant
Typical but non-limiting concentration be 1g/L, 2g/L, 3g/L, 4g/L, 5g/L, 5.5g/L, 6g/L, 7g/L, 8g/L, 9g/L,
9.5g/L、10g/L、11g/L、12g/L、14g/L、15g/L、16g/L、18g/L、20g/L、22g/L、24g/L、25g/L、26g/
L, 28g/L, 30g/L, 32g/L, 35g/L, 36g/L, 38g/L, 40g/L, 42g/L, 45g/L, 48g/L or 50g/L.
As further preferred technical solution, it includes following ingredient that the excrement, which saves liquid:
Chelating agent 20-300mmol/L, bacteriostatic agent 200-500mL/L, ionic strength maintain agent 5-10g/L and denaturant 5-
30g/L。
In the preferred embodiment of the present invention, by reasonably adjusting the proportion between each component, each component is given full play to
Between coordinated effect, more effectively inhibit DNA degradation and cell damage in fecal sample, maintain cellular morphology, suppression
Degrading enzyme processed and microbial activity better effect.
As further preferred technical solution, it includes following ingredient that the excrement, which saves liquid:
Chelating agent 30-100mmol/L, bacteriostatic agent 200-300mL/L, ionic strength maintain agent 8-10g/L and denaturant 5-
20g/L。
In the preferred embodiment of the present invention, by further adjusting the proportion between each component, each group is given full play to
/ coordinated effect, more effectively inhibit DNA degradation and cell damage in fecal sample, maintain cellular morphology,
Inhibit degrading enzyme and microbial activity better effect.
As further preferred technical solution, the pH value that the excrement saves liquid is 6.0-11.0；In the preferred reality
It applies in mode, by adjusting pH value in the range of 6.0-11.0, effectively excrement pH being maintained to stablize, maintaining cellular morphology, make
Obtain the DNA degradation not easy to break in fecal sample.
As further preferred technical solution, the pH value that the excrement saves liquid is 7.0-11.0；In the preferred reality
It applies in mode, by rationally adjusting pH in neutrality to alkaline range, effectively excrement pH is maintained to stablize, maintain cellular morphology, make
Obtain the DNA degradation not easy to break in fecal sample.
As further preferred technical solution, the pH value that the excrement saves liquid is 7.0-8.0；Preferably implement at this
In mode, by rationally adjusting within the scope of the neutrality to alkalescent that pH is 7.0-8.0, effectively excrement pH is maintained to stablize, remain thin
Born of the same parents' form, the DNA degradation not easy to break in sample.
As further preferred technical solution, the pH that the excrement saves liquid is mediated by the way that buffer is added；?
In the preferred embodiment, the buffer of addition offsets to a certain extent, mitigates remaining ingredient to the shadow of solution acid alkalinity
It rings, to keep the pH value of solution relatively stable.
As further preferred technical solution, the buffer includes that Tris-HCl, phosphate buffer, acetate are slow
Fliud flushing or Na2HPO4Sodium citrate buffer solution；In the preferred embodiment, pass through Tris-HCl, phosphate buffer, vinegar
Phthalate buffer or Na2HPO4- sodium citrate buffer solution keep the pH value of solution relatively stable.
As further preferred technical solution, the chelating agent includes organic sequestering agent；In the preferred embodiment
In, organic sequestering agent is influenced smaller by pH value, and sequestering power is more stable.
As further preferred technical solution, the organic sequestering agent includes edetate, 1,2- hexamethylene two
The double aminoethyl ether tetraacethyls of amine tetraacetate, diethyl pentetic acid salt, trientine salt, ethylene glycol, two sulphur of diethyl
For at least one of carbamic acid, nitrilotriacetic acid and ethylenediamine disuccinic acid trisodium.
As further preferred technical solution, the organic sequestering agent includes edetate and 1,2- hexamethylene two
At least one of amine tetraacetate；In the preferred embodiment, edetate and 1,2- cyclohexanediamine tetrem
Hydrochlorate is to Mg2+、Ca2+、Mn2+、Fe2+There is stronger chelation, the trace meter made in excrement will be chelated, thus
Inhibit the activity of enzyme.
As further preferred technical solution, the bacteriostatic agent includes substituted or unsubstituted C1-C4Alcohol, wherein described
Substituent group includes fluorine, chlorine or bromine；Typical but non-limiting substituted or unsubstituted C1-C4Alcohol be methanol, ethyl alcohol, 1- chlorethanol,
Ethylene glycol, 1- propyl alcohol, isopropanol, n-butyl alcohol, 2- butanol, 1,1- propylene glycol, 1,2-PD, 2,2- propylene glycol, 1,3- the third two
Alcohol, methaform or 1,1- butanediol etc..In the preferred embodiment, substituted or unsubstituted C1-C4Alcohol can be effectively suppressed micro-
Biological growth.
As further preferred technical solution, the bacteriostatic agent includes methanol, ethyl alcohol, ethylene glycol, isopropanol and neoprene
At least one of alcohol.
As further preferred technical solution, the bacteriostatic agent is at least one of ethyl alcohol and methaform；It is excellent at this
In the embodiment of choosing, microbial growth is can be effectively suppressed in ethyl alcohol and methaform.
As further preferred technical solution, it includes sodium chloride that the ionic strength, which maintains agent,；Preferably implement at this
In mode, sodium chloride can effectively maintain the stabilization of total ionic strength.
As further preferred technical solution, the denaturant includes anionic surfactant and amphoteric ion surface
At least one of activating agent；In the preferred embodiment, anionic surfactant can effectively will be in fecal sample
Nucleolysis enzyme such as protease, DNA enzymatic (DNases) and RNA enzyme (RNases) carry out denaturation treatment, while making microorganism not
It is easy to breed.
As further preferred technical solution, the anionic surfactant includes that sulfate type anionic surface is living
Property at least one of agent and sulfonate type anionic surfactant.
As further preferred technical solution, the sulfate type anionic surfactant includes urea, dodecane
Base sodium sulphate, ammonium lauryl sulfate, lithium dodecyl sulfate, sodium decyl sulfate, sodium octyl sulfate, ethoxylated dodecyl alcohol
Sodium sulfovinate, sulfated castor oil, lauryl sulfate diethanolamine, dodecyltriethanolamine sulfate, lauryl potassium sulfate, the moon
At least one of osmanthus base sulfuric acid triethanolamine, lauryl sulfate monoethanolamine and lauryl magnesium sulfate；Preferably implement at this
In mode, above-mentioned sulfate type anionic surfactant can be effectively by the nucleolysis enzyme in fecal sample such as protease, DNA
Enzyme (DNases) and RNA enzyme (RNases) carry out denaturation treatment, while microorganism being made to be not easy to breed.
As further preferred technical solution, the sulfate type anionic surfactant includes urea and dodecane
At least one of base sodium sulphate；In the preferred embodiment, urea or lauryl sodium sulfate can be effectively by excrement samples
Nucleolysis enzyme such as protease, DNA enzymatic (DNases) and RNA enzyme (RNases) in this carry out denaturation treatment, while making micro-
Biology is not easy to breed.
As further preferred technical solution, the sulfonate type anionic surfactant includes two isobutyl of succinic acid
Ester sodium sulfonate, sulfosuccinic acid -1,4- diamyl ester sodium, sulfo-succinic acid dihexyl sodium, sulfosuccinate dicyclohexyl maleate sodium, 11
In alkenyl single ethanol amide Disodium sulfosuccinate, sulfosuccinic acid lauryl disodium and Sodium docusate at least
It is a kind of；In the preferred embodiment, sulfonate type anionic surfactant can effectively drop the nucleic acid in fecal sample
It solves enzyme such as protease, DNA enzymatic (DNases) and RNA enzyme (RNases) carries out denaturation treatment, while microorganism being made to be not easy to grow
It is raw.
As further preferred technical solution, the sulfonate type anionic surfactant is that sulfosuccinic acid two is pungent
Ester sodium；In the preferred embodiment, Sodium docusate can effectively by the nucleolysis enzyme in fecal sample such as
Protease, DNA enzymatic (DNases) and RNA enzyme (RNases) carry out denaturation treatment, while microorganism being made to be not easy to breed.
As further preferred technical solution, the zwitterionic surfactant includes in chlorination guanidine and guanidine thiocyanate
At least one；In the preferred embodiment, chlorination guanidine or guanidine thiocyanate be may make the nucleolysis in fecal sample
Enzyme such as protease, DNA enzymatic (DNases) and RNA enzyme (RNases) carry out denaturation treatment, while microorganism being made to be not easy to breed.
As further preferred technical solution, the zwitterionic surfactant is guanidine thiocyanate.
According to the second aspect of the invention, the preparation method that the excrement saves liquid is provided, including by the original
Material is configured to solution, obtains excrement and saves liquid.
Excrement preservation liquid can be obtained by mixing raw material in the method for the present invention.Preparation method is simple, it is easy to operate, be easy to
Implement, raw material sources are wide, it is economical and easily available, be nontoxic, environment-friendly type raw material.The present invention is to environment, place, equipment etc. without special limit
System, used low in raw material price, safety and environmental protection performance is good, and low for equipment requirements, cost of investment is low, practicability and adaptability
It by force, is a kind of environmentally friendly, energy saving, efficiently, inexpensive excrement preservation liquid and preparation method thereof.
As further preferred technical solution, after raw material is configured to solution, is filtered using filter membrane device, obtain excrement
Save liquid；It in the preferred embodiment, is filtered by filter membrane device, isolates the impurity in excrement preservation liquid, it is filtered
Without chemical change in journey.
According to the third aspect of the present invention, it provides the excrement and saves application of the liquid in excrement preservation.
When excrement of the present invention saves liquid and saves fecal sample, at normal temperature can long-term preservation fecal sample, can effectively press down
DNA degradation and cell damage in fecal sample processed maintain cellular morphology, inhibit degrading enzyme and microbial activity in fecal sample.
According to the fourth aspect of the present invention, a kind of Preservation methods of faeces is provided, the Preservation methods of faeces is to use
The excrement saves liquid and saves.
Preservation methods of faeces of the present invention saves liquid by using excrement of the invention, can long-term preservation excrement sample at normal temperature
This.
It is stored in heparin tube it is understood that excrement can be saved liquid by us, convenient for users to carrying and transporting,
When in use, it is transferred out of excrement from heparin tube and saves liquid.Likewise, we can also will preserve the excrement of fecal sample
Just preservation liquid, which is placed in heparin tube, is saved.
Technical solution of the present invention is described further below in conjunction with embodiment and comparative example.
Embodiment 1
1, excrement saves liquid
A kind of excrement preservation liquid, it includes following ingredient that the excrement, which saves liquid:
Chelating agent is ethylenediamine tetra-acetic acid tripotassium, concentration 60mmol/L；Bacteriostatic agent is ethyl alcohol, concentration 230mL/L；From
It is sodium chloride, concentration 9g/L that sub- intensity, which maintains agent,；Denaturant is lauryl sodium sulfate, concentration 10g/L, and buffer is
Tris-HCl, the pH that excrement saves liquid is 7.5.
2, excrement saves the preparation of liquid
A kind of excrement saves the preparation method of liquid, comprising the following steps:
(1) respectively by the ethylenediamine tetra-acetic acid tripotassium of 60 molar parts, the ethyl alcohol of 230 parts by volume, 9 parts by weight sodium chloride,
The lauryl sodium sulfate of 10 parts by weight is dissolved in ultrapure water, is using the pH that buffer Tris-HCl adjusts excrement preservation liquid
7.5, after ultrapure water constant volume, the concentration that obtained excrement saves ethylenediamine tetra-acetic acid tripotassium in liquid is 300mmol/L, ethyl alcohol it is dense
Degree is 230mL/L, and the concentration of sodium chloride is 9g/L, and the concentration of lauryl sodium sulfate is 10g/L.
Wherein, the ratio of the molar part, parts by volume and parts by weight is mmol:mL:g.
(2) the preservation liquid obtained using disposable filter membrane device filtration step (1), is stored in liquid storage bottle.
Embodiment 2-7
Embodiment 2-7 difference from example 1 is that, obtain excrement save each ingredient in liquid concentration it is different, tool
Body is as shown in table 1.
The excrement of 1 embodiment 2-7 of table saves the concentration of each ingredient in liquid
Embodiment 8-11
Embodiment 8-11 difference from example 1 is that, obtain excrement save liquid pH it is different, specifically such as 2 institute of table
Show.
The excrement of 2 embodiment 8-11 of table saves the pH of liquid
| Embodiment 8 | Embodiment 9 | Embodiment 10 | Embodiment 11 | |
| pH | 6.0 | 11.0 | 7.0 | 8.0 |
Comparative example 1-9
Comparative example 1-9 difference from example 1 is that, obtain excrement save each ingredient in liquid concentration it is different, tool
Body is as shown in table 3.
The excrement of 3 comparative example 1-9 of table saves the concentration of each ingredient in liquid
| Chelating agent mmol/L | Bacteriostatic agent mL/L | Sodium chloride g/L | Denaturant g/L | |
| Comparative example 1 | 800 | — | — | — |
| Comparative example 2 | 5 | — | — | — |
| Comparative example 3 | 0 | — | — | — |
| Comparative example 4 | — | 40 | — | — |
| Comparative example 5 | — | 800 | — | — |
| Comparative example 6 | — | — | 0.2 | — |
| Comparative example 7 | — | — | 18 | — |
| Comparative example 8 | — | — | — | 0.2 |
| Comparative example 9 | — | — | — | 80 |
In table 3, "-" represents same as Example 1.
Comparative example 10
Comparative example 10 difference from example 1 is that, obtain excrement save liquid pH be 4.
Test example 1
At 25 DEG C, liquid is saved using the excrement that 1-11 of the embodiment of the present invention and comparative example 1-10 are obtained, 0 is saved to excrement
It, 3 days, 5 days and at 7 days, detect its DNA concentration, obtain that the results are shown in Table 4.
4 DNA concentration of table
In table 4, negative control group is physiological saline.
As shown in Table 4, the excrement of embodiment 1-11 saves liquid can save fecal sample for a long time at normal temperature, and can have
Effect inhibits DNA degradation and cell damage in fecal sample, maintains cellular morphology, inhibits degrading enzyme and microbial activity effect good,
The Sample preservation time is long.And DNA gradually degrades in comparative example 1-10 and negative control group (physiological saline), concentration decline.
Test example 2
The excrement preservation liquid and negative control group (physiological saline) preservation fecal sample being prepared using embodiment 1,7 days
After carry out electrophoresis experiment, obtain agarose gel electrophoresis figure as depicted in figs. 1 and 2, Fig. 1 is that 1 excrement of the embodiment of the present invention is protected
Liquid storage saves the agarose gel electrophoresis figure that 7 days fecal samples obtain, and is from left to right respectively to save the 0th day, the 3rd day, the 5th day
With the 7th day, Fig. 2 is that negative control group (physiological saline) saves the obtained agarose gel electrophoresis figure of 7 days fecal samples, from a left side to
The right side is respectively to save the 0th day, the 3rd day, the 5th day and the 7th day.
Genomic DNA after 1 excrement of embodiment preservation liquid, 7 days fecal samples of preservation it can be seen from Fig. 1 and Fig. 2 is complete,
And negative group DNA degrades.
Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；To the greatest extent
Pipe present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that: its according to
So be possible to modify the technical solutions described in the foregoing embodiments, or to some or all of the technical features into
Row equivalent replacement；And these are modified or replaceed, various embodiments of the present invention technology that it does not separate the essence of the corresponding technical solution
The range of scheme.
Claims (10)
1. a kind of excrement saves liquid, which is characterized in that it includes following ingredient that the excrement, which saves liquid:
Chelating agent 10-500mmol/L, bacteriostatic agent 100-600mL/L, ionic strength maintain agent 1-10g/L and denaturant 1-50g/
L。
2. excrement according to claim 1 saves liquid, which is characterized in that it includes following ingredient that the excrement, which saves liquid:
Chelating agent 20-300mmol/L, bacteriostatic agent 200-500mL/L, ionic strength maintain agent 5-10g/L and denaturant 5-30g/
L。
3. excrement according to claim 1 saves liquid, which is characterized in that it includes following ingredient that the excrement, which saves liquid:
Chelating agent 30-100mmol/L, bacteriostatic agent 200-300mL/L, ionic strength maintain agent 8-10g/L and denaturant 5-20g/
L。
4. excrement according to claim 1-3 saves liquid, which is characterized in that the pH value that the excrement saves liquid is
6.0-11.0 preferably 7.0-11.0, further preferably 7.0-8.0；
Preferably, the pH that the excrement saves liquid is mediated by the way that buffer is added；
Preferably, the buffer includes Tris-HCl, phosphate buffer, acetate buffer or Na2HPO4Sodium citrate
Buffer.
5. excrement according to claim 1-3 saves liquid, which is characterized in that the chelating agent includes organic chelated
Agent；
Preferably, the organic sequestering agent includes edetate, 1,2-cyclohexanediaminetetraacetic acid salt, diethyl triamine five
The double aminoethyl ether tetraacethyls of acetate, trientine salt, ethylene glycol, diethyldithiocar bamic acid, nitrilotriacetic acid
At least one of with ethylenediamine disuccinate；
Preferably, the organic sequestering agent includes at least one in edetate and 1,2-cyclohexanediaminetetraacetic acid salt
Kind.
6. excrement according to claim 1-3 saves liquid, which is characterized in that the bacteriostatic agent is including replacing or not
Substituted C1-C4Alcohol, wherein the substituent group includes fluorine, chlorine or bromine；
Preferably, the bacteriostatic agent includes at least one of methanol, ethyl alcohol, ethylene glycol, isopropanol and methaform, preferably second
At least one of pure and mild methaform；
Preferably, it includes sodium chloride that the ionic strength, which maintains agent,.
7. excrement according to claim 1-3 saves liquid, which is characterized in that the denaturant includes anion table
At least one of face activating agent and zwitterionic surfactant；
Preferably, the anionic surfactant includes sulfate type anionic surfactant and sulfonate type anion table
At least one of face activating agent；
Preferably, the sulfate type anionic surfactant includes urea, lauryl sodium sulfate, dodecyl sulphate
Ammonium, lithium dodecyl sulfate, sodium decyl sulfate, sodium octyl sulfate, ethoxylated dodecyl alcohol sodium sulfovinate, sulphation castor-oil plant
Oil, lauryl sulfate diethanolamine, dodecyltriethanolamine sulfate, lauryl potassium sulfate, triethanolamine lauryl sulfate, the moon
At least one of osmanthus base diethanolamine and lauryl magnesium sulfate, preferably in urea and lauryl sodium sulfate at least
It is a kind of；
Preferably, the sulfonate type anionic surfactant includes sulfosuccinic acid-diisobutylester-S-sodium salt, sulfosuccinic acid -1,
4- diamyl ester sodium, sulfo-succinic acid dihexyl sodium, sulfosuccinate dicyclohexyl maleate sodium, hendecene base single ethanol amide succinate
At least one of sodium sulfonate, sulfosuccinic acid lauryl disodium and Sodium docusate, preferably sulfosuccinic acid
Dioctyl ester sodium；
Preferably, the zwitterionic surfactant includes at least one of chlorination guanidine and guanidine thiocyanate, preferably sulphur cyanogen
Sour guanidine.
8. the preparation method that the described in any item excrement of claim 1-7 save liquid, which is characterized in that including matching the raw material
Solution is made, obtains excrement and saves liquid；
Preferably, it after raw material being configured to solution, is filtered using filter membrane device, obtains excrement and save liquid.
9. the described in any item excrement of claim 1-7 save application of the liquid in excrement preservation.
10. a kind of Preservation methods of faeces, which is characterized in that the Preservation methods of faeces is using any one of claim 1-7 institute
The excrement stated saves liquid and saves.
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