CN110286009A - A kind of mucous membrane of rectum cast-off cells sample pretreatment solution and preparation method thereof - Google Patents
A kind of mucous membrane of rectum cast-off cells sample pretreatment solution and preparation method thereof Download PDFInfo
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- CN110286009A CN110286009A CN201910491708.4A CN201910491708A CN110286009A CN 110286009 A CN110286009 A CN 110286009A CN 201910491708 A CN201910491708 A CN 201910491708A CN 110286009 A CN110286009 A CN 110286009A
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- rectum
- sample
- liquid
- cast
- mucous membrane
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- 210000004400 mucous membrane Anatomy 0.000 title claims abstract description 24
- 210000000664 rectum Anatomy 0.000 title claims abstract description 24
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- 239000007788 liquid Substances 0.000 claims abstract description 76
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 26
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 claims abstract description 23
- 229910052740 iodine Inorganic materials 0.000 claims abstract description 23
- 239000011630 iodine Substances 0.000 claims abstract description 23
- 229940100555 2-methyl-4-isothiazolin-3-one Drugs 0.000 claims abstract description 20
- BEGLCMHJXHIJLR-UHFFFAOYSA-N methylisothiazolinone Chemical compound CN1SC=CC1=O BEGLCMHJXHIJLR-UHFFFAOYSA-N 0.000 claims abstract description 20
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims abstract description 19
- BHNQPLPANNDEGL-UHFFFAOYSA-N 2-(4-octylphenoxy)ethanol Chemical compound CCCCCCCCC1=CC=C(OCCO)C=C1 BHNQPLPANNDEGL-UHFFFAOYSA-N 0.000 claims abstract description 14
- QCGGXGCODUUTLZ-UHFFFAOYSA-N [Na].[Na].[Na].[Na] Chemical compound [Na].[Na].[Na].[Na] QCGGXGCODUUTLZ-UHFFFAOYSA-N 0.000 claims abstract description 14
- 239000011780 sodium chloride Substances 0.000 claims abstract description 13
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims abstract description 12
- 229910052708 sodium Inorganic materials 0.000 claims abstract description 12
- 239000011734 sodium Substances 0.000 claims abstract description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- 239000012153 distilled water Substances 0.000 claims description 12
- 239000011159 matrix material Substances 0.000 claims description 3
- 239000002202 Polyethylene glycol Substances 0.000 claims description 2
- -1 ethyl Phenyl Chemical group 0.000 claims description 2
- 238000012856 packing Methods 0.000 claims description 2
- 229920001223 polyethylene glycol Polymers 0.000 claims description 2
- 238000002203 pretreatment Methods 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 11
- 238000001514 detection method Methods 0.000 abstract description 9
- 238000000034 method Methods 0.000 abstract description 8
- PNDPGZBMCMUPRI-UHFFFAOYSA-N iodine Chemical compound II PNDPGZBMCMUPRI-UHFFFAOYSA-N 0.000 abstract description 5
- 244000005700 microbiome Species 0.000 abstract description 5
- 230000008569 process Effects 0.000 abstract description 4
- 210000003097 mucus Anatomy 0.000 abstract description 3
- 230000001376 precipitating effect Effects 0.000 abstract description 3
- 210000004027 cell Anatomy 0.000 description 27
- 239000000243 solution Substances 0.000 description 20
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical group O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 12
- 230000006870 function Effects 0.000 description 9
- 239000003795 chemical substances by application Substances 0.000 description 8
- 230000008030 elimination Effects 0.000 description 7
- 238000003379 elimination reaction Methods 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 229920001661 Chitosan Polymers 0.000 description 5
- 241000233866 Fungi Species 0.000 description 5
- 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 5
- 229930182566 Gentamicin Natural products 0.000 description 5
- 229960002518 gentamicin Drugs 0.000 description 5
- 230000000007 visual effect Effects 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 210000003743 erythrocyte Anatomy 0.000 description 4
- 238000007689 inspection Methods 0.000 description 4
- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical compound [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 description 3
- 239000007983 Tris buffer Substances 0.000 description 3
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 239000008139 complexing agent Substances 0.000 description 3
- 238000004090 dissolution Methods 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 239000003755 preservative agent Substances 0.000 description 3
- 230000002335 preservative effect Effects 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- 102000002322 Egg Proteins Human genes 0.000 description 2
- 108010000912 Egg Proteins Proteins 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 230000000845 anti-microbial effect Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000003139 buffering effect Effects 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 210000001072 colon Anatomy 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 239000003317 industrial substance Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
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- 231100000252 nontoxic Toxicity 0.000 description 2
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- 230000007170 pathology Effects 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 239000013049 sediment Substances 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- 229910006299 γ-FeOOH Inorganic materials 0.000 description 2
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- 241000224489 Amoeba Species 0.000 description 1
- 206010003694 Atrophy Diseases 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- QXNVGIXVLWOKEQ-UHFFFAOYSA-N Disodium Chemical class [Na][Na] QXNVGIXVLWOKEQ-UHFFFAOYSA-N 0.000 description 1
- 239000003109 Disodium ethylene diamine tetraacetate Substances 0.000 description 1
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 description 1
- 108010034145 Helminth Proteins Proteins 0.000 description 1
- 241000607764 Shigella dysenteriae Species 0.000 description 1
- 208000037386 Typhoid Diseases 0.000 description 1
- 230000002924 anti-infective effect Effects 0.000 description 1
- 239000002260 anti-inflammatory agent Substances 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000001741 anti-phlogistic effect Effects 0.000 description 1
- 230000002421 anti-septic effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000037444 atrophy Effects 0.000 description 1
- 230000003385 bacteriostatic effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000003399 chemotactic effect Effects 0.000 description 1
- 238000005660 chlorination reaction Methods 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 235000019301 disodium ethylene diamine tetraacetate Nutrition 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 201000003373 familial cold autoinflammatory syndrome 3 Diseases 0.000 description 1
- 229910001448 ferrous ion Inorganic materials 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000010794 food waste Substances 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 244000000013 helminth Species 0.000 description 1
- 238000000703 high-speed centrifugation Methods 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
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- 230000001900 immune effect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 description 1
- 239000000693 micelle Substances 0.000 description 1
- 239000003094 microcapsule Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000002541 mycostatic Substances 0.000 description 1
- 230000001557 mycostatic effect Effects 0.000 description 1
- 210000004681 ovum Anatomy 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000000242 pagocytic effect Effects 0.000 description 1
- 230000003071 parasitic effect Effects 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 235000004252 protein component Nutrition 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 201000001275 rectum cancer Diseases 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
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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
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/30—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests characterised by the surfactants
-
- 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
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/14—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
- A01N43/16—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings with oxygen as the ring hetero atom
-
- 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
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/72—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with nitrogen atoms and oxygen or sulfur atoms as ring hetero atoms
- A01N43/80—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with nitrogen atoms and oxygen or sulfur atoms as ring hetero atoms five-membered rings with one nitrogen atom and either one oxygen atom or one sulfur atom in positions 1,2
-
- 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
- A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
- A01N59/12—Iodine, e.g. iodophors; Compounds thereof
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
Abstract
The present invention relates to a kind of mucous membrane of rectum cast-off cells sample pretreatment solution, the group for including is divided into methylisothiazolinone, shell iodine, trishydroxymethylaminomethane, disodium salt disodium, Nonidet P40, sodium chloride.The invention further relates to the preparation methods of above-mentioned pretreatment liquid.The present invention can provide stable environment to cell sample, and having prevents tangible mass sets and precipitating in sample, remove the mucus in sample, reduce the effect of viscosity；Because of the use in conjunction of shell iodine and methylisothiazolinone, has the function of dissipation to the microorganism especially mould in mucous membrane of rectum cast-off cells pattern detection pretreatment process and inhibit to breed；Because of the use in conjunction of isotonic sodium chlorrde solution, trishydroxymethylaminomethane, disodium salt disodium, Nonidet P40, the integrality and good form of mucous membrane of rectum cast-off cells can be maintained, to guarantee the accuracy of cell sample detection.
Description
Technical field
The invention belongs to medical supplies field, in particular to a kind of mucous membrane of rectum cast-off cells sample pretreatment solution and its system
Make method.
Background technique
Exfoliative cytology belongs to a branch of cell pathology, is the epithelial cell for acquiring partes corporis humani position, through dyeing
Afterwards with its form of micro- sem observation, a Men Xueke of clinical diagnosis disease is assisted.The advantages of exfoliative cytology checks has: simple easy
Row, high safety not high to equipment requirement；The pain caused by patient is few, and can draw materials inspection repeatedly；Diagnosis is rapid, cancer cell
Recall rate is higher, the follow-up observation especially suitable for extensive Cancer screen and people at highest risk.The inspection of mucous membrane of rectum exfoliative cytology
Looking into is one of the important means of carcinoma of the rectum screening.But rectal contents are more complicated, the liquid of digestive secretion containing enteral, pathology
Property diffusate, residual food waste, cast-off cells, bacterium, mould, helminth etc., may be there are also remaining excrement, especially loose stools.
Intrarectal microorganism forms certain interference to the preservation and detection of sample, so, mucous membrane of rectum exfoliative cytology inspection needs
Pre-treatment is carried out to sample.In pathologic sample, it is also possible to have pathogenic microbes, as shigella dysenteriae, typhoid bacillus,
Amoeba, parasitic ovum etc. make sample after detection have the potential danger of pollution.
Pretreatment liquid is the component part of external diagnosis reagent, needs certain validity period, generally 1~2 year.Liquid-based examination
Often there is fungus growth in agent management process, causes the filiform for occurring the formation such as fungal hyphae and spore in reagent, floccule
Even block.Fungus growth will seriously affect the accuracy of testing result.In order to solve the above problem, it needs a kind of with dissipation
Function, remove sample in microorganism, have effects that anticorrosion sterilizing, inhibit fungus growth, improve analysis detection precision and
The preconditioning technique of reliability.
Summary of the invention
The first object of the present invention be in order to solve the above technical problems, providing a kind of mucous membrane of rectum cast-off cells sample before
Manage liquid.The pretreatment liquid can rapidly process the sample of separate sources, complete the inspection of mucous membrane of rectum exfoliative cytology, improve detection
Precision.
The second object of the present invention is to provide the preparation method of above-mentioned mucous membrane of rectum cast-off cells sample pretreatment solution.
The present invention is achieved through the following technical solutions:
One, a kind of mucous membrane of rectum cast-off cells sample pretreatment solution, the component and content for including in every liter of solution are methyl
Isothiazolinone 0.1ml, shell iodine 65g, trishydroxymethylaminomethane 72.8g, disodium salt disodium 9.3g, ethylphenyl are poly-
Ethylene glycol 0.2g.
Specifically, the mucous membrane of rectum cast-off cells sample pretreatment solution, the matrix of the pretreatment liquid is isotonic chlorination
Sodium solution.
Two, the preparation method of above-mentioned mucous membrane of rectum cast-off cells sample pretreatment solution, the specific steps are as follows:
Isotonic sodium chlorrde solution is made Step 1: sodium chloride is dissolved in distilled water；
Step 2: trishydroxymethylaminomethane, disodium salt disodium, Nonidet P40 are dissolved in step 1
In solution obtained；
It is mixed Step 3: shell iodine and methylisothiazolinone are dissolved in liquid made from step 2 again.
Step 4: packing, completion.
Mechanism of the invention and and each industrial chemicals function and effect in the present invention it is as follows:
Methylisothiazolinone (MIT) is a kind of sterilization antiseptic of wide spectrum, can effectively kill algae, bacterium and fungi.
The activity single dose effective dose is few, nontoxic and pollution-free, is easily blended in all kinds of formulas, and PH use scope is wide, and dilution uses dense
After degree, it is easy to be biodegradable as nontoxic and pollution-free substance.Toxicity is low, noresidue is discharged, with various emulsifiers, surface-active
Agent and protein component compatibility are good.
Shell iodine (chitosan-iodo-complexes) has both the performance of chitosan and iodine.Chitosan in vivo can chemotactic leucocyte, lure
Local macrophage is led, its phagocytic function and hydrolysing activity are enhanced, stimulates it to generate lymphokine and inflammatory mediator, to enhance
The anti-infection ability of body promotes the healing of wound, has the characteristics such as immunological regulation, antibacterial, antitumor.Chitosan molecule H branch
The combination of the H chain of chain and iodine molecule, and the positive negative electricity of substance is attracting, thus iodine molecule by chitosan molecule be encapsulated as nanoscale capsule and
The form of micro-capsule.When effect, nanoscale iodine molecule Continuous slow release from chitosan molecule comes out bactericidal antiphlogistic, is able to maintain iodine and holds
Continuous 72 hours antibacterial and anti-inflammation functions have broad spectrum antimicrobial efficiency, have special efficacy to mould.Nano grade biological soldier takes
The shell iodine factor and the clinical chemical synthesis iodine of external application it is different: the latter is inorganic iodine, is strong oxidizer, is usually used in skin surface and glues
Film disinfection, only the dissipation effect of short time.The former is the molecular structure phase of biological active iodine and the self anti-inflammatory cells of human body
Seemingly, has efficient antimicrobial and anti-inflammation efficacy.
Trishydroxymethylaminomethane (Tris) provides weakly alkaline environment for the pretreatment liquid of the present invention, its pKa is 8.1；
Theoretical according to buffering, effective buffering range of Tris buffer is between pH7.0 to 9.2.Tris also has surfactant
Effect.
Nonidet P40 is the detergent without ion, and critical micelle concentration is low, strong with protein binding power, is used for
Prevent material molecule hydrophobic interaction, it is ensured that the abundant dissolution of albumen and stable structure are generally usually used in various cells, egg
White, groupization etc. operation, particularly for memebrane protein it is non denatured under the conditions of dissolution.
Disodium ethylene diamine tetraacetate is a kind of complexing agent, is mainly used for that ferrous ion is complexed, controls polymerization rate.Benefit
With the dispersion of complexing agent, suspension function, the generation of sediment in water is prevented.Complexing agent and Fe2+The complex of formation can absorb portion
Divide visible light, generates the free radical of Strong oxdiative type, make initial reaction stage Fe2+Quickly it is oxidized to Fe3+, fast oxidation rate is conducive to life
At γ-FeOOH (γ-FeOOH) of low degree of crystallization, tangible mass sets and precipitating in sample are also prevented, and has and removes
Mucus in sample reduces the function of viscosity.
The matrix of the pretreatment liquid is isotonic sodium chlorrde solution, is isostension solution, has the integrality of protection cell
Function.
Good effect by adopting the above technical scheme: one kind provided by the invention has the function of that dissipation mucous membrane of rectum falls off carefully
Born of the same parents' sample pretreatment solution can provide stable environment to cell sample；With preventing tangible mass sets and precipitating in sample,
The mucus in sample is removed, the effect of viscosity is reduced；In addition, because of the use in conjunction of shell iodine and methylisothiazolinone, to straight
Microorganism especially mould in intestinal mucosa cast-off cells pattern detection pretreatment process has the function of dissipation and inhibits to breed；
Because isotonic sodium chlorrde solution, trishydroxymethylaminomethane, disodium salt disodium, combining for Nonidet P40 answer
With the integrality of mucous membrane of rectum cast-off cells can be maintained；To guarantee the accuracy of cell sample detection.
Specific embodiment
The present invention is described further combined with specific embodiments below, as described below, is only to of the invention preferable
Embodiment not limits the present invention, and any person skilled in the art is possibly also with above-mentioned
The technology contents of announcement are changed or are modified as the equivalent embodiment changed on an equal basis.Without departing from the concept of the present invention,
Any simple modification, equivalent change and modification made to the above embodiment according to the technical essence of the invention all fall within this hair
In bright protection scope.
Embodiment 1
Embodiment 1 illustrates mould bacteriostatic test
One, the pretreatment liquid of different elimination agents is configured
1, liquid 1 (pretreatment liquid of the present invention) is verified
(1) 9g sodium chloride is dissolved in distilled water 1000ml；
(2) trishydroxymethylaminomethane 72.8g, disodium salt disodium 9.3g, Nonidet P40 0.2g
It is dissolved in upper liquid；
(3) shell iodine 65g and methylisothiazolinone 0.1ml are dissolved in aforesaid liquid and are mixed again.
2, liquid 2 (containing only elimination agent) is verified
(1) 9g sodium chloride is dissolved in distilled water 1000ml；
(2) shell iodine 65g and methylisothiazolinone 0.1ml are dissolved in aforesaid liquid and are mixed.
3, liquid 3 (no elimination agent) is verified
(1) 9g sodium chloride is dissolved in distilled water 1000ml；
(2) trishydroxymethylaminomethane 72.8g, disodium salt disodium 9.3g, Nonidet P40 0.2g
It is dissolved in upper liquid.
4, it verifies liquid 4 (containing only isotonic sodium chlorrde solution): 9g sodium chloride being dissolved in distilled water 1000ml and is dissolved.
5, verify liquid 5 (elimination agent is formaldehyde)
(1) 9g sodium chloride is dissolved in distilled water 1000ml；
(2) trishydroxymethylaminomethane 72.8g, disodium salt disodium 9.3g, Nonidet P40 0.2g
It is dissolved in upper liquid；
(3) formaldehyde 10ml is dissolved in aforesaid liquid again and is mixed.
6, verify liquid 6 (elimination agent is gentamicin)
(1) 9g sodium chloride is dissolved in distilled water 1000ml；
(2) trishydroxymethylaminomethane 72.8g, disodium salt disodium 9.3g, Nonidet P40 0.2g
It is dissolved in upper liquid；
(3) 12 unit of gentamicin is dissolved in aforesaid liquid again and is mixed.
Two, sample liquid is configured
1, sample collection: selecting suitable cell sampler, gently wipes in colon interior position, obtains mucous membrane of rectum and falls off
Cell sample.
2, preparation method: 6 parts of swabs after sampling are inserted into verifying each 2ml of liquid 1-6 respectively and are gently agitated for, made on swab
The sample and verifying liquid of acquisition mix, and sample liquid 1-6 is made respectively.
Three, mycotic culture test method:
1, each 2ml of this liquid 1-6 is sampled with aseptic operation to be added separately in plate, use 25ml, 45-47 DEG C of Meng in time
Add and draw red agar pour plate, rotation plate is uniformly mixed.Agar solidification, positive horizontalization plate.
2, it cultivates five days for 28 degree Celsius
3, plate mold colony number is counted respectively.
Four, test result such as table 1
1 sample liquid 1-6 fungus growth situation of table
1, the pretreatment liquid (sample liquid 1) of the present invention, containing only the pretreatment liquid (sample liquid 2) of elimination agent, to be showed no mould raw
It is long；
2, pretreatment liquid (sample liquid 3-4) the mould sprawling growth without any effective preservative covers entire plate, note
Record is that bacterium colony is spread.
3, it is 33CFU that preservative, which is pretreatment liquid (sample liquid 5) clump count of formaldehyde,.
4, it is 17CFU that preservative, which is pretreatment liquid (sample liquid 6) clump count of gentamicin,.
5, statistical procedures are shown, iodine containing shell and methylisothiazolinone with without shell iodine and methylisothiazolinone before
Treatment fluid compares, iodine containing shell and methylisothiazolinone are compared with formaldehyde or gentamicin, and P value is respectively less than 0.01, has very
Significant difference.
Five, conclusion: iodine containing shell and methylisothiazolinone mucous membrane of rectum cast-off cells Sample pretreatment provided by the invention
Liquid can effectively kill mould.Compared with formaldehyde and gentamicin, shell iodine has stronger with methylisothiazolinone use in conjunction
Dissipation and inhibit mould breed the effect of.
Embodiment 2
Embodiment 2 illustrates that cell integrity is observed
One, the pretreatment liquid of differential tension is configured
1, liquid 1 (pretreatment liquid of the present invention) is verified
(1) 9g sodium chloride is dissolved in distilled water 1000ml；
(2) trishydroxymethylaminomethane 72.8g, disodium salt disodium 9.3g, Nonidet P40 0.2g
It is dissolved in upper liquid；
(3) shell iodine 65g and methylisothiazolinone 0.1ml are dissolved in aforesaid liquid and are mixed again.
2, liquid 2 (low-tension pretreatment liquid) is verified
(1) distilled water 1000ml is taken；
(2) trishydroxymethylaminomethane 72.8g, disodium salt disodium 9.3g, Nonidet P40 0.2g
It is dissolved in upper liquid；
(3) shell iodine 65g and methylisothiazolinone 0.1ml are dissolved in aforesaid liquid and are mixed again.
3, liquid 3 (high-tension pretreatment liquid) is verified
(1) sodium chloride is incorporated into distilled water 1000ml to dissolution saturation state (sodium chloride about 360g)；
(2) trishydroxymethylaminomethane 72.8g, disodium salt disodium 9.3g, Nonidet P40 0.2g
It is dissolved in upper liquid；
(3) shell iodine 65g and methylisothiazolinone 0.1ml are dissolved in aforesaid liquid and are mixed again.
4, it verifies liquid 4: 9g sodium chloride is dissolved in distilled water 1000ml, isotonic sodium chlorrde solution is made.
Two, sample liquid is configured
1, sample collection: selecting suitable cell swab, gently wipes in colon interior position, obtains mucous membrane of rectum and falls off carefully
Born of the same parents' sample.
2, preparation method: swab is inserted into respectively in verifying each 5ml of liquid 1-4 after sampling and stirs mixing, sample liquid 1- is made
4.Every part of sample liquid with pipettor accurately to draw each 20 microlitres uniform stall with goods spread out on the ground for sale cloth of sediment therein after 3000 turns of high speed centrifugations
In on the flat surface in glass slide center, covered.
Three, test result
1, cell integrity observation method (being calculated with red blood cell):
(1) the standard visual field is set: glass slide being placed on objective table, is placed on the light bar hole of eyepiece with piece, then observes.
It is tangent at a distance of two parallel lines of 1.382mm and the visual field on glass slide；Four side of block plaid with piece (mircrometer gauge) also with visual field phase
It cuts.
(3) high power microscopic observation red blood cell.Each smear checks 10 visuals field, and 10 visuals field checked will be equably
It is distributed on measurement room, can be controlled with the propeller for having scale on microscope carrier, from top to bottom, or from left to right one
Every trade is regularly observed.Complete mean constant of red blood cell is calculated with counter.
2, test result such as table 2:
2 intact red blood cells number of table
It is statistically analyzed as the result is shown:
(1) compared with low-tension pretreatment liquid (sample liquid 2), P value has very isostension (sample liquid 1) less than 0.01
Significant difference；
(2) for isostension (sample liquid 1) compared with high-tension pretreatment liquid (sample liquid 3), P value is greater than 0.05, and there is no aobvious
Write difference.But high power microscopic observation red cell morphology, high-tension pretreatment liquid cellular atrophy wrinkle is flat, loses normal morphology.
Simple isotonic sodium chlorrde solution (sample liquid 4) uses trishydroxymethylaminomethane, disodium salt two with merging
The industrial chemicals such as sodium, Nonidet P40 and shell iodine and methylisothiazolinone elimination agent pretreatment liquid (sample liquid 1) ratio
Compared with P value is greater than 0.05, and significant difference is not present.But simple isotonic sodium chlorrde solution does not have the mycostatic function of suppression,
Through illustrating in test example 1.
The above test is not as it can be seen that simple isotonic sodium chlorrde solution and hypertonic liquid have dissipation and keep cell integrity
And the double effects of good form.
In conclusion mucous membrane of rectum cast-off cells provided by the invention detect dedicated pretreatment liquid, can effectively kill mould
Microorganisms such as bacterium, and can guarantee the integrality and good form of sample inner cell, exclude sample memory various factors it is dry
It disturbs, improves the precision of detection, be a kind of for detecting the suitable pretreatment liquid of mucous membrane of rectum cast-off cells.
Claims (3)
1. a kind of mucous membrane of rectum cast-off cells sample pretreatment solution, it is characterised in that: the component and content for including in every liter of solution
For methylisothiazolinone 0.1ml, shell iodine 65g, trishydroxymethylaminomethane 72.8g, disodium salt disodium 9.3g, ethyl
Phenyl polyethylene glycol 0.2g.
2. a kind of mucous membrane of rectum cast-off cells sample pretreatment solution according to claim 1, it is characterised in that: the pre-treatment
The matrix of liquid is isotonic sodium chlorrde solution.
3. a kind of preparation method of mucous membrane of rectum cast-off cells sample pretreatment solution according to claim 1 or 2, feature
Be: specific step is as follows:
Step 1: sodium chloride is dissolved in distilled water；
Step 2: trishydroxymethylaminomethane, disodium salt disodium, Nonidet P40 are dissolved in step 1 and are made
Solution in；
It is mixed Step 3: shell iodine and methylisothiazolinone are dissolved in liquid made from step 2 again.
Step 4: packing, completion.
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