CN110283726A - A kind of liquid basal cell and microbiological treatment save reagent and preparation method thereof - Google Patents
A kind of liquid basal cell and microbiological treatment save reagent and preparation method thereof Download PDFInfo
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- CN110283726A CN110283726A CN201910595692.1A CN201910595692A CN110283726A CN 110283726 A CN110283726 A CN 110283726A CN 201910595692 A CN201910595692 A CN 201910595692A CN 110283726 A CN110283726 A CN 110283726A
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- Prior art keywords
- reagent
- basal cell
- microbiological treatment
- liquid basal
- save reagent
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- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 54
- 239000007788 liquid Substances 0.000 title claims abstract description 40
- 210000000270 basal cell Anatomy 0.000 title claims abstract description 39
- 230000002906 microbiologic effect Effects 0.000 title claims abstract description 38
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims abstract description 21
- 239000000203 mixture Substances 0.000 claims abstract description 15
- 239000003963 antioxidant agent Substances 0.000 claims abstract description 14
- 230000003078 antioxidant effect Effects 0.000 claims abstract description 13
- 239000000049 pigment Substances 0.000 claims abstract description 13
- 239000004909 Moisturizer Substances 0.000 claims abstract description 11
- 230000001333 moisturizer Effects 0.000 claims abstract description 11
- 230000003204 osmotic effect Effects 0.000 claims abstract description 11
- 239000006174 pH buffer Substances 0.000 claims abstract description 11
- 239000004094 surface-active agent Substances 0.000 claims abstract description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 9
- 239000008367 deionised water Substances 0.000 claims abstract description 7
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 7
- 239000000686 essence Substances 0.000 claims abstract description 7
- 239000002994 raw material Substances 0.000 claims description 18
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid group Chemical group C(CC(O)(C(=O)O)CC(=O)O)(=O)O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 15
- 238000012545 processing Methods 0.000 claims description 14
- 238000010907 mechanical stirring Methods 0.000 claims description 13
- 238000000034 method Methods 0.000 claims description 11
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 9
- UUIVKBHZENILKB-UHFFFAOYSA-N 2,2-dibromo-2-cyanoacetamide Chemical compound NC(=O)C(Br)(Br)C#N UUIVKBHZENILKB-UHFFFAOYSA-N 0.000 claims description 7
- 239000007853 buffer solution Substances 0.000 claims description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical group OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 6
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 6
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical group [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 150000008442 polyphenolic compounds Chemical class 0.000 claims description 5
- 235000013824 polyphenols Nutrition 0.000 claims description 5
- RZSYLLSAWYUBPE-UHFFFAOYSA-L Fast green FCF Chemical compound [Na+].[Na+].C=1C=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C(=CC(O)=CC=2)S([O-])(=O)=O)C=CC=1N(CC)CC1=CC=CC(S([O-])(=O)=O)=C1 RZSYLLSAWYUBPE-UHFFFAOYSA-L 0.000 claims description 4
- 241001122767 Theaceae Species 0.000 claims description 4
- 229940107698 malachite green Drugs 0.000 claims description 4
- FDZZZRQASAIRJF-UHFFFAOYSA-M malachite green Chemical compound [Cl-].C1=CC(N(C)C)=CC=C1C(C=1C=CC=CC=1)=C1C=CC(=[N+](C)C)C=C1 FDZZZRQASAIRJF-UHFFFAOYSA-M 0.000 claims description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 3
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical group [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 claims description 3
- 239000008351 acetate buffer Substances 0.000 claims description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 3
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 3
- CZBZUDVBLSSABA-UHFFFAOYSA-N butylated hydroxyanisole Chemical compound COC1=CC=C(O)C(C(C)(C)C)=C1.COC1=CC=C(O)C=C1C(C)(C)C CZBZUDVBLSSABA-UHFFFAOYSA-N 0.000 claims description 3
- 239000008103 glucose Substances 0.000 claims description 3
- 235000011187 glycerol Nutrition 0.000 claims description 3
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 3
- 229920000053 polysorbate 80 Polymers 0.000 claims description 3
- 239000001103 potassium chloride Substances 0.000 claims description 3
- 235000011164 potassium chloride Nutrition 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- YBCVMFKXIKNREZ-UHFFFAOYSA-N acoh acetic acid Chemical compound CC(O)=O.CC(O)=O YBCVMFKXIKNREZ-UHFFFAOYSA-N 0.000 claims description 2
- 238000004090 dissolution Methods 0.000 claims 1
- 239000000463 material Substances 0.000 claims 1
- 239000000834 fixative Substances 0.000 abstract description 8
- 102000002322 Egg Proteins Human genes 0.000 abstract description 7
- 108010000912 Egg Proteins Proteins 0.000 abstract description 7
- 210000004681 ovum Anatomy 0.000 abstract description 7
- LEQAOMBKQFMDFZ-UHFFFAOYSA-N glyoxal Chemical compound O=CC=O LEQAOMBKQFMDFZ-UHFFFAOYSA-N 0.000 abstract description 6
- 238000003756 stirring Methods 0.000 abstract description 6
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 abstract description 3
- 239000004372 Polyvinyl alcohol Substances 0.000 abstract description 3
- 150000001299 aldehydes Chemical class 0.000 abstract description 3
- 239000012472 biological sample Substances 0.000 abstract description 3
- 229940015043 glyoxal Drugs 0.000 abstract description 3
- 229920002451 polyvinyl alcohol Polymers 0.000 abstract description 3
- 210000003812 trophozoite Anatomy 0.000 abstract description 3
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 abstract description 2
- 230000003071 parasitic effect Effects 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 29
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 19
- 239000000523 sample Substances 0.000 description 14
- 238000004321 preservation Methods 0.000 description 13
- 235000006708 antioxidants Nutrition 0.000 description 10
- 244000005700 microbiome Species 0.000 description 10
- 238000001514 detection method Methods 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- DLFVBJFMPXGRIB-UHFFFAOYSA-N Acetamide Chemical compound CC(N)=O DLFVBJFMPXGRIB-UHFFFAOYSA-N 0.000 description 4
- 238000003745 diagnosis Methods 0.000 description 4
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 4
- 229910052753 mercury Inorganic materials 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 235000013601 eggs Nutrition 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000007781 pre-processing Methods 0.000 description 3
- 239000004255 Butylated hydroxyanisole Substances 0.000 description 2
- 241000244203 Caenorhabditis elegans Species 0.000 description 2
- 208000030852 Parasitic disease Diseases 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 210000001124 body fluid Anatomy 0.000 description 2
- 239000010839 body fluid Substances 0.000 description 2
- 230000031709 bromination Effects 0.000 description 2
- 238000005893 bromination reaction Methods 0.000 description 2
- 229940043253 butylated hydroxyanisole Drugs 0.000 description 2
- 235000019282 butylated hydroxyanisole Nutrition 0.000 description 2
- 230000030833 cell death Effects 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 230000034994 death Effects 0.000 description 2
- 238000004043 dyeing Methods 0.000 description 2
- 230000000855 fungicidal effect Effects 0.000 description 2
- 239000000417 fungicide Substances 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 230000001590 oxidative effect Effects 0.000 description 2
- 230000000750 progressive effect Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 241000191940 Staphylococcus Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- RCTYPNKXASFOBE-UHFFFAOYSA-M chloromercury Chemical compound [Hg]Cl RCTYPNKXASFOBE-UHFFFAOYSA-M 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002550 fecal effect Effects 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- JMANVNJQNLATNU-UHFFFAOYSA-N glycolonitrile Natural products N#CC#N JMANVNJQNLATNU-UHFFFAOYSA-N 0.000 description 1
- 230000005802 health problem Effects 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 210000000697 sensory organ Anatomy 0.000 description 1
- BHZOKUMUHVTPBX-UHFFFAOYSA-M sodium acetic acid acetate Chemical compound [Na+].CC(O)=O.CC([O-])=O BHZOKUMUHVTPBX-UHFFFAOYSA-M 0.000 description 1
- 210000004215 spore Anatomy 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- 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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Dentistry (AREA)
- Environmental Sciences (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of liquid basal cells and microbiological treatment to save reagent and preparation method thereof, by the way that successively Halocyanoacetamide, moisturizer, pH buffer, osmotic pressure regulator, surfactant, antioxidant and pigment, essence are mixed evenly, and it adds mixture into the deionized water for being 30 DEG C~35 DEG C to water temperature and stirs, adding sodium hydroxide tune pH value is 5.5~6.5, and a kind of liquid basal cell disclosed by the invention can be obtained and microbiological treatment saves reagent.The reagent is the fixative for fixing biological sample, and the reagent is not mercurous and formaldehyde, can be compatible from different parasitic stages (such as sporangiocyst, ovum, larva and trophozoite), it is effective in the case where no addition polyvinyl alcohol or addition aldehyde (such as glyoxal, glutaraldehyde).
Description
Technical field
The invention belongs to technical field of medical detection, specially a kind of fixative more particularly to one of fixed biological sample
Kind liquid basal cell and microbiological treatment save reagent and preparation method thereof.
Background technique
Various types sample, especially to the detection of cell and the visible components such as microorganism in body fluid sample require into
The certain pre-processing process of row, this pre-processing can improve positive rate, and impurity is to visible component in reduction sample
Final detection, serious person cause detection invalid or can not carry out detection, it is seen then that in body fluid sample cell and microorganism etc.
Visible component detection carries out correct pre-processing and is of great significance for accuracy and the reliability etc. of testing result.Generally
For, in order to effectively detect cell or microorganism, need to save sample, transport, cracking, extracting and developing, precipitating,
The processing links such as fixation, film-making, dyeing.
And parasitic infection causes the with a long history of disease in human and animal, usually passes through dyeing and microscopic examination
Worm egg and larva or protozoan trophozoite and/or tumour in fecal specimens confirm the diagnosis of enterozoon, but no matter
Using which kind of method, a problem for diagnosing parasitic infection is to collect and check the delay between sample, if without suitable
Fixative, then some special enzymes that sample can be made to contain by itself or because breeding bacterium and micro- in environment present in sample
Biology and lead to sample fast degradation, and if do not handled in time, will affect the Accurate Diagnosis of enterozoon, and quasi-
The disconnected best fixation for depending on obtaining sample when collecting is made a definite diagnosis, therefore, fixed solution is commonly used in processing sample to be posted
Infested diagnosis.
Currently, the fixative for saving stool sample usually contains mercury, formaldehyde or formalin, with many limitations
The shortcomings that it is used.For example, the fixative based on mercury and the Schaudinn fixative containing mercury chloride alkali be although capable of providing it is good
Good definition, but due to cost is excessively high and is ready to handle the scarcity of the disposition company of mercury waste and there are handling problems, and
Preservative containing formaldehyde or formalin in fixative can cause similar environment and health problem.
Therefore, how to develop a kind of not mercurous or formaldehyde and there is biocompatibility, safely and effectively liquid basal cell and micro-
Biological treatment saves the problem of reagent is those skilled in the art's urgent need to resolve.
Summary of the invention
In view of this, being provided a kind of suitable for fixed life the purpose of the present invention is aiming at the problems existing in the prior art
The liquid basal cell and microbiological treatment of object sample save reagent, and disclose above-mentioned liquid basal cell and microbiological treatment preservation reagent
Preparation method.
To achieve the goals above, the present invention adopts the following technical scheme:
A kind of liquid basal cell and microbiological treatment save reagent, and the processing preservation reagent is to be prepared from the following raw materials, and press
Mass percent meter:
Preferably, it is to be prepared from the following raw materials that the processing, which saves reagent, by mass percentage:
Preferably, it is to be prepared from the following raw materials that the processing, which saves reagent, by mass percentage:
A kind of liquid basal cell is mixed with out by the coordinated of above-mentioned raw materials and microbiological treatment saves reagent, it should
Reagent is fixative for fixing biological sample, and the reagent is not mercurous and formaldehyde, can be from different parasitic stages (such as spores
Capsule, ovum, larva and trophozoite) it is compatible, be in the case where no addition polyvinyl alcohol or addition aldehyde (such as glyoxal, glutaraldehyde)
Effectively.
Preferably, the Halocyanoacetamide includes 2,2-Dibromo-2-cyanoacetamide, the bromo- 3- cyano third of 2,2- bis-
One of amide or multiple combinations.
It should be noted that the preferred Halocyanoacetamide of the present invention is 2,2-Dibromo-2-cyanoacetamide, and 2,2- bis-
Bromo- 2- cyanoacetamide is a kind of fungicide of broad-spectrum high efficacy, can readily penetrate through the cell membrane of microorganism, and is acted on certain
Albumen group restores the normal oxidation of cell and stops, so as to cause cell death.Meanwhile its being also an option that property of branch
Bromination or oxidizing microorganisms specific enzymes metabolin, eventually lead to microorganism death.
Preferably, the pH buffer is acetic acid-acetate buffer system, phosphoric acid buffer system or citric acid buffer body
System, and the moisturizer is glycerine, ethylene glycol or sorbierite.
The preferred pH buffer of the present invention is citric acid buffer system, and limit the pH range of the citric acid buffer system as
3~6.
Preferably, the surfactant is dodecyl sodium sulfate, Tween-80 or CO-40；And the antioxidant is
Tea polyphenols, butylated hydroxy anisole or 4,4- methylene-bis(2,6-di-t-butyl phenol).
Preferred anti-oxidant of the present invention is tea polyphenols, and tea polyphenols also have bacteriostasis in addition to antioxidation,
Such as there is inhibiting effect to staphylococcus, Escherichia coli and hay bacillus.
Preferably, the osmotic pressure regulator is sodium chloride, potassium chloride or glucose；And the pigment be malachite green,
One of fast green or multiple combinations.
The present invention is disclosed using the mixture of malachite green and fast green composition as pigment, can be formed in aqueous solution
A kind of bottle green, the bottle green can postpone the decomposition that 2,2-Dibromo-2-cyanoacetamide generates under conditions of having illumination, extend
The service life of this preservation liquid, and the pigment mixed system is formed by bottle green also and can improve the vision of human faecal mass in the solution
Sense organ.
It is a further aim of the invention provide it is a kind of easy to operate, it is not high to equipment requirement, be suitable for industrialized production
Liquid basal cell and microbiological treatment save reagent preparation method.
A kind of preparation method of liquid basal cell and microbiological treatment preservation reagent, specifically comprises the following steps:
(1) raw material is weighed respectively according to said ratio, pH buffer and antioxidant are add to deionized water, it is mechanical
It stirs to dissolve and mixed liquor is heated to 30 DEG C~35 DEG C；Halocyanoacetamide is then added, mechanical stirring keeps its molten
Solution；Then moisturizer, osmotic pressure regulator, surfactant and pigment, essence are sequentially added, mechanical stirring keeps all components molten
It solves and is uniformly mixed, obtain mixture I；
It should be noted that since Halocyanoacetamide is slightly soluble in water, and its aqueous solution easily decomposes under alkaline condition,
It is stable in acid condition.PH buffer and antioxidant is added first to provide a stable environment, then adds halogeno-cyanogen
Yl acetamide.
(2) adding sodium hydroxide tune pH value is 5.5~6.5 in the mixture I for obtaining step (1), and the present invention can be obtained
A kind of disclosed liquid basal cell and microbiological treatment save reagent.
As can be seen from the above technical solutions, compared with prior art, the present invention provides a kind of liquid basal cell and microorganisms
Processing saves reagent and preparation method thereof, has following excellent effect:
Firstly, not containing mercury or formaldehyde in raw material disclosed by the invention, application range is not only widened, but also there are currently no add in fact
Still effective purpose in the case where Polyvinyl alcohol or aldehyde (glyoxal or glutaraldehyde)；
Then, the preferred Halocyanoacetamide of the present invention is 2,2-Dibromo-2-cyanoacetamide, and 2,2- bis- bromo- 2- cyanogen
Yl acetamide is a kind of fungicide of broad-spectrum high efficacy, can readily penetrate through the cell membrane of microorganism, and acts on certain albumen collection
Group restores the normal oxidation of cell and stops, so as to cause cell death.Meanwhile the bromination of its being also an option that property of branch
Or the specific enzymes metabolin of oxidizing microorganisms, eventually lead to microorganism death；
Finally, preparation method disclosed by the invention is simple, and it is not high to equipment requirement, it is suitable for industrialized production.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this
The embodiment of invention for those of ordinary skill in the art without creative efforts, can also basis
The attached drawing of offer obtains other attached drawings.
Fig. 1 is the microscope figure of whipworm ovum of the present invention.
Fig. 2 is the microscope figure of roundworm fertilized eggs of the present invention.
Specific embodiment
Technical solution disclosed by the invention is clearly and completely described below in conjunction with the embodiment of the present invention, it is clear that
The described embodiment is only a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention,
Every other embodiment obtained by those of ordinary skill in the art without making creative efforts, belongs to this hair
The range of bright protection.
The embodiment of the invention discloses a kind of not mercurous or formaldehyde and there is biocompatibility, safely and effectively liquid basal cell
Reagent is saved with microbiological treatment, and specifically discloses the preparation method of the liquid basal cell and microbiological treatment preservation reagent, not only
It is easy to operate, it is also not high to equipment requirement, it is suitable for industrialized production.
For a better understanding of the invention, further specific elaboration is made to the present invention below by following embodiment, but not
It can be regarded as limitation of the invention, it is some nonessential according to made by foregoing invention content for those skilled in the art
It improves and adjusts, be also considered as and be within the scope of the present invention.
Saving reagent the invention discloses a kind of processing is to be prepared from the following raw materials, by mass percentage:
In order to further optimize the above technical scheme, Halocyanoacetamide includes 2,2-Dibromo-2-cyanoacetamide, 2,
One of bromo- 3- cyano propionamide of 2- bis- or multiple combinations.
In order to further optimize the above technical scheme, pH buffer is citric acid buffer system, phosphoric acid buffer system or second
Acid-acetate buffer system, and the moisturizer is glycerine, ethylene glycol or sorbierite.
In order to further optimize the above technical scheme, surfactant is dodecyl sodium sulfate, Tween-80 or CO-40；
And the antioxidant is tea polyphenols, butylated hydroxy anisole or 4,4- methylene-bis(2,6-di-t-butyl phenol).
In order to further optimize the above technical scheme, osmotic pressure regulator is sodium chloride, potassium chloride or glucose；And it is described
Pigment is one of malachite green, fast green or multiple combinations.
The invention also discloses the preparation methods that a kind of liquid basal cell and microbiological treatment save reagent, specifically include as follows
Step:
(1) raw material is weighed respectively according to said ratio, pH buffer and antioxidant are add to deionized water, it is mechanical
It stirs to dissolve and mixed liquor is heated to 30 DEG C~35 DEG C；Halocyanoacetamide is then added, mechanical stirring keeps its molten
Solution；Then moisturizer, osmotic pressure regulator, surfactant and pigment, essence are sequentially added, mechanical stirring keeps all components molten
It solves and is uniformly mixed, obtain mixture I；
(2) adding sodium hydroxide tune pH value is 5.5~6.5 in the mixture I for obtaining step (1), and the present invention can be obtained
A kind of disclosed liquid basal cell and microbiological treatment save reagent.
Technical solution disclosed by the invention is further described below in conjunction with specific embodiment.
Embodiment 1:
A kind of liquid basal cell and microbiological treatment save reagent, and the processing preservation reagent is to be prepared from the following raw materials, and press
Mass percent meter:
Embodiment 2:
A kind of liquid basal cell and microbiological treatment save reagent, and the processing preservation reagent is to be prepared from the following raw materials, and press
Mass percent meter:
Embodiment 3:
A kind of liquid basal cell and microbiological treatment save reagent, and the processing preservation reagent is to be prepared from the following raw materials, and press
Mass percent meter:
Embodiment 4:
A kind of liquid basal cell and microbiological treatment save reagent, and the processing preservation reagent is to be prepared from the following raw materials, and press
Mass percent meter:
Embodiment 5:
A kind of preparation method of liquid basal cell and microbiological treatment preservation reagent, the method specifically comprise the following steps:
(1) raw material is weighed respectively according to said ratio, pH buffer and antioxidant are add to deionized water, it is mechanical
It stirs to dissolve and mixed liquor is heated to 30 DEG C~35 DEG C；Halocyanoacetamide is then added, mechanical stirring keeps its molten
Solution；Then moisturizer, osmotic pressure regulator, surfactant and pigment, essence are sequentially added, mechanical stirring keeps all components molten
It solves and is uniformly mixed, obtain mixture I；
(2) adding sodium hydroxide tune pH value is 5.5 in the mixture I for obtaining step (1), be can be obtained disclosed by the invention
A kind of liquid basal cell and microbiological treatment save reagent.
Embodiment 6:
A kind of preparation method of liquid basal cell and microbiological treatment preservation reagent, the method specifically comprise the following steps:
(1) raw material is weighed respectively according to said ratio, pH buffer and antioxidant are add to deionized water, it is mechanical
It stirs to dissolve and mixed liquor is heated to 30 DEG C~35 DEG C；Halocyanoacetamide is then added, mechanical stirring keeps its molten
Solution；Then moisturizer, osmotic pressure regulator, surfactant and pigment, essence are sequentially added, mechanical stirring keeps all components molten
It solves and is uniformly mixed, obtain mixture I；
(2) adding sodium hydroxide tune pH value is 6.0 in the mixture I for obtaining step (1), be can be obtained disclosed by the invention
A kind of liquid basal cell and microbiological treatment save reagent.
Embodiment 7:
A kind of preparation method of liquid basal cell and microbiological treatment preservation reagent, the method specifically comprise the following steps:
(1) raw material is weighed respectively according to said ratio, pH buffer and antioxidant are add to deionized water, it is mechanical
It stirs to dissolve and mixed liquor is heated to 30 DEG C~35 DEG C；Halocyanoacetamide is then added, mechanical stirring keeps its molten
Solution；Then moisturizer, osmotic pressure regulator, surfactant and pigment, essence are sequentially added, mechanical stirring keeps all components molten
It solves and is uniformly mixed, obtain mixture I；
(2) adding sodium hydroxide tune pH value is 6.5 in the mixture I for obtaining step (1), be can be obtained disclosed by the invention
A kind of liquid basal cell and microbiological treatment save reagent.
Each embodiment in this specification is described in a progressive manner, the highlights of each of the examples are with other
The difference of embodiment, the same or similar parts in each embodiment may refer to each other.
The content of present invention is not limited only to the content of the various embodiments described above, and the combination of one of them or several embodiments is equally
The object of the invention may be implemented.
In order to further verify excellent effect of the invention, inventor has also carried out following experiment:
Experiment one:
For applicant to the comparative tests of different antioxidant additive amounts, specific experiment and interpretation of result are as follows:
| Control group | 1 | 2 | 3 | |
| Halocyanoacetamide | 2% | 2% | 2% | 2% |
| Moisturizer | 5% | 5% | 5% | 5% |
| PH buffer | 0.5% | 0.5% | 0.5% | 0.5% |
| Osmotic pressure regulator | 0.1% | 0.1% | 0.1% | 0.1% |
| Surfactant | 0.015% | 0.015% | 0.015% | 0.015% |
| Pigment | 0 | 0 | 0 | 0 |
| Essence | 0 | 0 | 0 | 0 |
| Antioxidant | 0 | 0.03% | 0.1% | 0.5% |
Above four groups of solution is placed in 25 DEG C of environment and is protected from light storage, control group at first week solution gradually by colourless
Solution becomes yellow solution, and experimental group 1 about started to become yellow solution from colourless solution when third week, and experimental group 2 is about
Started to become yellow solution from colourless solution at the 5th week, experimental group 3 places 2 months solution without significant change.
Above four groups of solution is placed in 25 DEG C of natural environment and is stored, control group is in third day solution gradually by by colourless
Solution becomes yellow solution, and experimental group 1 about started to become yellow solution from colourless solution when first week, and experimental group 2 is about
Start to become yellow solution from colourless solution in second week, experimental group 3 about starts to become yellow from colourless solution in 4th week
Solution.
Experiment two:
The liquid basal cell and microbiological treatment guarantor that the present invention refers to are placed in the fresh excreta that sample spoon takes 5g5 years old child
It deposits in reagent, collects worm's ovum using Kato- Katz technique, and observe under the microscope, specific experiment interpretation of result is as follows:
It is learnt by Fig. 1 and Fig. 2, the integrality of worm's ovum structure is presented in whipworm ovum and roundworm fertilized eggs under the microscope,
It is able to illustrate that the present invention discloses the liquid basal cell of preparation and microbiological treatment saves reagent and has good preservation effect to worm's ovum.
Each embodiment in this specification is described in a progressive manner, the highlights of each of the examples are with other
The difference of embodiment, the same or similar parts in each embodiment may refer to each other.For method disclosed in embodiment
For, since it is corresponded to the methods disclosed in the examples, so being described relatively simple, related place is said referring to method part
It is bright.
The foregoing description of the disclosed embodiments enables those skilled in the art to implement or use the present invention.
Various modifications to these embodiments will be readily apparent to those skilled in the art, as defined herein
General Principle can be realized in other embodiments without departing from the spirit or scope of the present invention.Therefore, of the invention
It is not intended to be limited to the embodiments shown herein, and is to fit to and the principles and novel features disclosed herein phase one
The widest scope of cause.
Claims (8)
1. a kind of liquid basal cell and microbiological treatment save reagent, which is characterized in that it is by following original that the processing, which saves reagent,
Material is made, by mass percentage:
2. a kind of liquid basal cell according to claim 1 and microbiological treatment save reagent, which is characterized in that the processing
Saving reagent is to be prepared from the following raw materials, by mass percentage:
3. a kind of liquid basal cell according to claim 1 and microbiological treatment save reagent, which is characterized in that the processing
Saving reagent is to be prepared from the following raw materials, by mass percentage:
4. a kind of any described liquid basal cell and microbiological treatment save reagent according to claim 1~3, which is characterized in that
The Halocyanoacetamide includes one of 2,2- dibromo-2-cyanoacetamide, the bromo- 3- cyano propionamide of 2,2- bis- or more
Kind combination.
5. a kind of any described liquid basal cell and microbiological treatment save reagent according to claim 1~3, which is characterized in that
The pH buffer is citric acid buffer system, phosphoric acid buffer system or acetic acid-acetate buffer system, and the moisturizer is
Glycerine, ethylene glycol or sorbierite.
6. a kind of any described liquid basal cell and microbiological treatment save reagent according to claim 1~3, which is characterized in that
The surfactant is dodecyl sodium sulfate, Tween-80 or CO-40；And the antioxidant is tea polyphenols, butylhydroxy
Anisole or 4,4- methylene-bis(2,6-di-t-butyl phenol).
7. a kind of any described liquid basal cell and microbiological treatment save reagent according to claim 1~3, which is characterized in that
The osmotic pressure regulator is sodium chloride, potassium chloride or glucose；And the pigment is one of malachite green, fast green or more
Kind combination.
8. the preparation method that a kind of liquid basal cell as described in claim 1 and microbiological treatment save reagent, which is characterized in that
The method specifically comprises the following steps:
(1) raw material is weighed respectively according to said ratio, pH buffer and antioxidant are add to deionized water, mechanical stirring
It makes it dissolve and mixed liquor is heated to 30 DEG C~35 DEG C；Halocyanoacetamide is then added, mechanical stirring makes it dissolve；So
After sequentially add moisturizer, osmotic pressure regulator, surfactant and pigment, essence, mechanical stirring makes all components dissolution simultaneously
It is uniformly mixed, obtains mixture I；
(2) adding sodium hydroxide tune pH value is 5.5~6.5 in the mixture I for obtaining step (1), and the present invention can be obtained and disclose
A kind of liquid basal cell and microbiological treatment save reagent.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110973117A (en) * | 2019-12-23 | 2020-04-10 | 苏州堪赛尔医学检验有限公司 | Tissue cell fixing solution and preparation method and application method thereof |
| CN111057645A (en) * | 2019-12-31 | 2020-04-24 | 百沃星联(上海)环保科技有限公司 | Microbial storage device for food decomposition and storage method thereof |
| CN113907064A (en) * | 2021-10-08 | 2022-01-11 | 广州达安基因股份有限公司 | Liquid-based cell preservation solution and preparation method thereof |
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| CN103688152A (en) * | 2011-05-02 | 2014-03-26 | 乔斯·卡洛斯·拉潘那 | Fixative solution, for fixation and preservation of biological samples |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110973117A (en) * | 2019-12-23 | 2020-04-10 | 苏州堪赛尔医学检验有限公司 | Tissue cell fixing solution and preparation method and application method thereof |
| CN110973117B (en) * | 2019-12-23 | 2021-11-23 | 苏州堪赛尔医学检验有限公司 | Rat eyeball histiocyte fixing solution as well as preparation method and application method thereof |
| CN111057645A (en) * | 2019-12-31 | 2020-04-24 | 百沃星联(上海)环保科技有限公司 | Microbial storage device for food decomposition and storage method thereof |
| CN111057645B (en) * | 2019-12-31 | 2023-11-14 | 百沃星联(上海)环保科技有限公司 | Microbial storage device for food decomposition and storage method thereof |
| CN113907064A (en) * | 2021-10-08 | 2022-01-11 | 广州达安基因股份有限公司 | Liquid-based cell preservation solution and preparation method thereof |
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