CN107991151A - A kind of reagent for hemocyte analyzers - Google Patents
A kind of reagent for hemocyte analyzers Download PDFInfo
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- CN107991151A CN107991151A CN201711170124.4A CN201711170124A CN107991151A CN 107991151 A CN107991151 A CN 107991151A CN 201711170124 A CN201711170124 A CN 201711170124A CN 107991151 A CN107991151 A CN 107991151A
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- reagent
- hemolytic agents
- sodium
- hemocyte analyzers
- hemolytic
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- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 74
- 210000003677 hemocyte Anatomy 0.000 title claims abstract description 22
- 229940000351 hemocyte Drugs 0.000 title claims abstract description 22
- 239000003219 hemolytic agent Substances 0.000 claims abstract description 58
- 239000000243 solution Substances 0.000 claims abstract description 32
- 238000010790 dilution Methods 0.000 claims abstract description 21
- 239000012895 dilution Substances 0.000 claims abstract description 21
- 238000004043 dyeing Methods 0.000 claims abstract description 21
- 238000004140 cleaning Methods 0.000 claims abstract description 19
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 52
- 238000003756 stirring Methods 0.000 claims description 31
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 30
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 29
- 239000008367 deionised water Substances 0.000 claims description 28
- 229910021641 deionized water Inorganic materials 0.000 claims description 28
- 239000011780 sodium chloride Substances 0.000 claims description 26
- 230000003204 osmotic effect Effects 0.000 claims description 21
- FJKROLUGYXJWQN-UHFFFAOYSA-N 4-hydroxybenzoic acid Chemical compound OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 claims description 18
- 238000010438 heat treatment Methods 0.000 claims description 15
- 229910052938 sodium sulfate Inorganic materials 0.000 claims description 15
- 235000011152 sodium sulphate Nutrition 0.000 claims description 15
- 238000001914 filtration Methods 0.000 claims description 13
- 239000007983 Tris buffer Substances 0.000 claims description 9
- 150000002460 imidazoles Chemical class 0.000 claims description 9
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims description 9
- 229940090248 4-hydroxybenzoic acid Drugs 0.000 claims description 8
- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 claims description 8
- HXWGXXDEYMNGCT-UHFFFAOYSA-M decyl(trimethyl)azanium;chloride Chemical compound [Cl-].CCCCCCCCCC[N+](C)(C)C HXWGXXDEYMNGCT-UHFFFAOYSA-M 0.000 claims description 7
- DDXLVDQZPFLQMZ-UHFFFAOYSA-M dodecyl(trimethyl)azanium;chloride Chemical compound [Cl-].CCCCCCCCCCCC[N+](C)(C)C DDXLVDQZPFLQMZ-UHFFFAOYSA-M 0.000 claims description 7
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 6
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 claims description 6
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 6
- IVKNZCBNXPYYKL-UHFFFAOYSA-N 2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethanol Chemical compound CC(C)(C)CC(C)(C)C1=CC=C(OCCOCCOCCOCCOCCOCCOCCOCCOCCOCCO)C=C1 IVKNZCBNXPYYKL-UHFFFAOYSA-N 0.000 claims description 5
- 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 claims description 5
- DWCZIOOZPIDHAB-UHFFFAOYSA-L methyl green Chemical compound [Cl-].[Cl-].C1=CC(N(C)C)=CC=C1C(C=1C=CC(=CC=1)[N+](C)(C)C)=C1C=CC(=[N+](C)C)C=C1 DWCZIOOZPIDHAB-UHFFFAOYSA-L 0.000 claims description 5
- 239000003755 preservative agent Substances 0.000 claims description 5
- 230000002335 preservative effect Effects 0.000 claims description 5
- IZUPJOYPPLEPGM-UHFFFAOYSA-M sodium;hydron;phthalate Chemical compound [Na+].OC(=O)C1=CC=CC=C1C([O-])=O IZUPJOYPPLEPGM-UHFFFAOYSA-M 0.000 claims description 5
- 229920001030 Polyethylene Glycol 4000 Polymers 0.000 claims description 4
- 229940125717 barbiturate Drugs 0.000 claims description 4
- WWAABJGNHFGXSJ-UHFFFAOYSA-N chlorophenol red Chemical compound C1=C(Cl)C(O)=CC=C1C1(C=2C=C(Cl)C(O)=CC=2)C2=CC=CC=C2S(=O)(=O)O1 WWAABJGNHFGXSJ-UHFFFAOYSA-N 0.000 claims description 4
- SFNALCNOMXIBKG-UHFFFAOYSA-N ethylene glycol monododecyl ether Chemical compound CCCCCCCCCCCCOCCO SFNALCNOMXIBKG-UHFFFAOYSA-N 0.000 claims description 4
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 3
- 239000004471 Glycine Substances 0.000 claims description 3
- 235000019253 formic acid Nutrition 0.000 claims description 3
- 238000012417 linear regression Methods 0.000 abstract description 12
- 238000004519 manufacturing process Methods 0.000 abstract description 5
- 238000003912 environmental pollution Methods 0.000 abstract description 2
- 231100000252 nontoxic Toxicity 0.000 abstract description 2
- 230000003000 nontoxic effect Effects 0.000 abstract description 2
- 239000002994 raw material Substances 0.000 abstract description 2
- 238000012360 testing method Methods 0.000 description 25
- 238000000034 method Methods 0.000 description 11
- 210000003743 erythrocyte Anatomy 0.000 description 10
- 210000004369 blood Anatomy 0.000 description 8
- 239000008280 blood Substances 0.000 description 8
- 210000000601 blood cell Anatomy 0.000 description 8
- 238000004458 analytical method Methods 0.000 description 7
- 239000003795 chemical substances by application Substances 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 210000004027 cell Anatomy 0.000 description 6
- 238000001816 cooling Methods 0.000 description 6
- 210000000265 leukocyte Anatomy 0.000 description 6
- 238000011156 evaluation Methods 0.000 description 5
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 210000003714 granulocyte Anatomy 0.000 description 4
- 210000001772 blood platelet Anatomy 0.000 description 3
- 210000004698 lymphocyte Anatomy 0.000 description 3
- ONJQDTZCDSESIW-UHFFFAOYSA-N polidocanol Chemical compound CCCCCCCCCCCCOCCOCCOCCOCCOCCOCCOCCOCCOCCO ONJQDTZCDSESIW-UHFFFAOYSA-N 0.000 description 3
- 229910052708 sodium Inorganic materials 0.000 description 3
- 239000011734 sodium Substances 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- SNRUBQQJIBEYMU-UHFFFAOYSA-N Dodecane Natural products CCCCCCCCCCCC SNRUBQQJIBEYMU-UHFFFAOYSA-N 0.000 description 2
- 102000001554 Hemoglobins Human genes 0.000 description 2
- 108010054147 Hemoglobins Proteins 0.000 description 2
- 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 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 125000000217 alkyl group Chemical group 0.000 description 2
- AJXBTRZGLDTSST-UHFFFAOYSA-N amino 2-methylprop-2-enoate Chemical compound CC(=C)C(=O)ON AJXBTRZGLDTSST-UHFFFAOYSA-N 0.000 description 2
- 235000019270 ammonium chloride Nutrition 0.000 description 2
- -1 barbiturates Chemical compound 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 125000003438 dodecyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000004065 semiconductor Substances 0.000 description 2
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical compound CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 description 1
- 102000010445 Lactoferrin Human genes 0.000 description 1
- 108010063045 Lactoferrin Proteins 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 230000005465 channeling Effects 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 238000005660 chlorination reaction Methods 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 238000002242 deionisation method Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 210000003979 eosinophil Anatomy 0.000 description 1
- CSSYQJWUGATIHM-IKGCZBKSSA-N l-phenylalanyl-l-lysyl-l-cysteinyl-l-arginyl-l-arginyl-l-tryptophyl-l-glutaminyl-l-tryptophyl-l-arginyl-l-methionyl-l-lysyl-l-lysyl-l-leucylglycyl-l-alanyl-l-prolyl-l-seryl-l-isoleucyl-l-threonyl-l-cysteinyl-l-valyl-l-arginyl-l-arginyl-l-alanyl-l-phenylal Chemical compound C([C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 CSSYQJWUGATIHM-IKGCZBKSSA-N 0.000 description 1
- 229940078795 lactoferrin Drugs 0.000 description 1
- 235000021242 lactoferrin Nutrition 0.000 description 1
- 238000002356 laser light scattering Methods 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 208000010110 spontaneous platelet aggregation Diseases 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
Classifications
-
- 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
-
- 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
- G01N1/34—Purifying; Cleaning
-
- 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
- G01N2001/302—Stain compositions
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a kind of reagent for hemocyte analyzers, it includes dilution, LEOI hemolytic agents, LEOII dyeing liquors, LH hemolytic agents, LBA hemolytic agents, cleaning solution.The reagent for hemocyte analyzers of the present invention can step the replacement reagent of auspicious 5800 original-pack reagents of BC as the classification cytoanalyze of the prior art five, the problem of its is of high cost, delivery cycle is long can be efficiently solved, and with genuine reagent linear regression coefficient R up to more than 0.9900, system has preferable correlation；The raw material sources of the present invention are extensive, are easy to get, and cheap, nontoxic, environmental pollution is few；The simple production process of product of the present invention, is easy to duplication of production, and stability is good, long shelf-life, is worth further genralrlization.
Description
Technical field
The present invention relates to medical diagnosis instrument reagent, and in particular to a kind of reagent for hemocyte analyzers.
Background technology
Mindray is the producer of the domestic classification cellanalyzer of production five of having the ability earliest, and wherein BC-5800 is one
There is money binary channels leucocyte five to classify, and DIFF passages can dissolve red blood cell and blood platelet in DIFF laser channelings, and to thermophilic
Eosinophil carries out specific stain, by laser light scattering and FCM analysis, makes lymphocyte, monocyte, acidophilus
Property granulocyte, neutrophil cell distinguish to come completely, while screen abnormal lymphocytes (ALY) and huge immature cell
(LIC), it is shown on DIFF scatter diagrams.In addition, having an independent BASO passage, the effect through too strong molten type reagent is realized molten
The specific stain of blood and basophilic granulocyte, can effectively exclude slightly solubility red blood cell, Giant platelet, platelet aggregation and
Influence of the factors such as other acellular particles to counting, realizes the accurate detection of basophilic granulocyte and total white blood cells.
Under semiconductor laser scattering and cell dyeing technical role, with reference to holographic digital processing technology, to Heterotypic Lymphocyte
Cell (ALY) and huge immature cell (LIC) carry out quantitative detection, improve laboratory abnormal cell is screened it is reliable
Property.
But the original package reagent consumption of auspicious BC-5800 cellanalyzers advanced in years and demand are all very big at present, and
The cost and price of reagent is expensive, causes medical treatment cost height.
The content of the invention
In view of the problems of the existing technology, the present invention provides a kind of reagent for hemocyte analyzers, particularly step auspicious
BC-5800 cellanalyzer replacement reagents.
The present invention uses following technical scheme：
A kind of reagent for hemocyte analyzers, it include dilution, LEOI hemolytic agents, LEOII dyeing liquors, LH hemolytic agents,
LBA hemolytic agents, cleaning solution；
The dilution includes 2.5~8.0g/L of sodium chloride, 3.5~8.0g/L of sodium sulphate, 0.5~5.0g/L of imidazoles, sweet
0.1~3.0g/L of propylhomoserin, 0.3~1.0g/L of EDTA-2Na, 0.3~1.0g/L of preservative；
The LEOI hemolytic agents include 1.0~6.0g/L of decyl trimethyl ammonium chloride, 1.0~4.0g/L of OP-10,
1.0~6.0g/L of PEG4000,2.0~10.0g/L of P-hydroxybenzoic acid, 1.0~10.0g/L of Trisbase, sodium chloride 2.0
~10.0g/L, 2~5.0g/L of sodium sulphate, 1.0~3.0g/L of methyl green；
The LEOII dyeing liquors include 1.0~4.0g/L of dodecyl trimethyl ammonium chloride, 0.5~4.0g/ of TX-100
L, 0.5~5.0g/L of P-hydroxybenzoic acid, 2.0~5.0g/L of hydrogen phthalate sodium, 1.0~5.0g/L of sodium chloride；
The LH hemolytic agents include 5.0~20.0g/L of decyl trimethyl ammonium chloride, 1.0~5.0g/L of Brij35, first
1.0~8.0g/L of acid, 10.0~25.0g/L of sodium chloride；
The LBA hemolytic agents include 2.0~10.0g/L of dodecyl trimethyl ammonium chloride, 0.5~3.0g/ of barbiturates
L, 1.0~10.0g/L of Trisbase, 10.0~20.0g/L of sodium chloride, 5~10mg/L of chlorophenol red；
The cleaning solution include 0.5~10.0g/L of PE6800,0.5~10.0g/L of AEO-9, sodium chloride 1.0~
10.0g/L, 1.0~10.0g/L of sodium sulphate, 0.5~5.0g/L of imidazoles, 0.1~1.0g/L of sodium azide.
Further, the pH value of the dilution is 6.5~7.0, and the electrical conductivity of the dilution is 14.0~17.0ms/
Cm, the osmotic pressure of the dilution is 260~320mOsm/kg.
Further, sequentially added in 1000mL beakers the imidazoles weighed, glycine, sodium chloride, sodium sulphate,
EDTA-2Na, adds suitable deionized water, and heating stirring is cooled to less than 40 DEG C, addition has weighed up after it is completely dissolved
Preservative, stir evenly, with the volume of deionized water constant volume to 1L, measure the pH value, electrical conductivity, osmotic pressure of solution, filtering
Afterwards up to the dilution.
Further, the pH value of the LEOI hemolytic agents is 5.50~6.0, and the electrical conductivity of the LEOI hemolytic agents is 9.0
~10.0ms/cm, the osmotic pressure of the LEOI hemolytic agents is 200~250mOsm/kg.
Further, sequentially added in 1000mL beakers the decyl trimethyl ammonium chloride weighed, OP-10,
PEG4000, Trisbase, P-hydroxybenzoic acid, sodium chloride, sodium sulphate, add suitable deionized water, and heating stirring treats that its is complete
After fully dissolved, add methyl green, with the volume of deionized water constant volume to 1L, stir evenly, measure the pH value of solution, electrical conductivity,
Osmotic pressure, up to the LEOI hemolytic agents after filtering.
Further, the pH value of the LEOII dyeing liquors is 5.50~6.0, and the electrical conductivity of the LEOII dyeing liquors is
6.0~7.0ms/cm, the osmotic pressure of the LEOII dyeing liquors is 100~150mOsm/kg.
Further, the dodecyl trimethyl ammonium chloride weighed, TX-100, right is sequentially added in 1000mL beakers
Hydroxybenzoic acid, hydrogen phthalate sodium, sodium chloride, sodium sulphate, add suitable deionized water, and heating stirring makes it completely molten
Solution, with the volume of deionized water constant volume to 1L, stirs evenly, measures the pH value, electrical conductivity, osmotic pressure of solution, after filtering to obtain the final product
The LEOII dyeing liquors.
Further, the pH value of the LH hemolytic agents is 2.0~3.0, the electrical conductivity of the LH hemolytic agents for 35.0~
40.0ms/cm, the osmotic pressure of the LH hemolytic agents is 700~800mOsm/kg.
Further, the decyl trimethyl ammonium chloride weighed, Brij35, chlorination are sequentially added in 1000mL beakers
Sodium, adds suitable deionized water, and heating stirring is completely dissolved it, after being cooled to 40 DEG C, adds the formic acid weighed up, stirs
Mix, add remaining deionized water constant volume to the volume of 1L, stir evenly, measure the pH value, electrical conductivity, osmotic pressure of solution, mistake
Up to LH hemolytic agents after filter.
Further, the pH value of the LBA hemolytic agents is 9.0~11.0, the electrical conductivity of the LBA hemolytic agents for 13.0~
16.0ms/cm, the osmotic pressure of the LBA hemolytic agents is 400~500mOsm/kg.
Further, sequentially added in 1000mL beakers the dodecyl trimethyl ammonium chloride weighed, barbiturates,
Trisbase, sodium chloride, add suitable deionized water, and heating stirring is completely dissolved it, and after cooling, it is molten to add chlorophenol red
Solution, adds remaining deionized water, the volume of constant volume to 1L, stirs evenly, and measures the pH value, electrical conductivity, osmotic pressure of solution, mistake
Up to LBA hemolytic agents after filter.
Further, the pH value of the cleaning solution is 8.0~10.0, the electrical conductivity of the cleaning solution for 15.0~
20.0ms/cm。
Further, the PE6800, AEO-9 weighed, imidazoles, sodium chloride, sulfuric acid are sequentially added in 1000mL beakers
Sodium, adds suitable deionized water, and heating stirring is completely dissolved it, and sodium azide dissolving is added after cooling, is added remaining
Deionized water, the volume of constant volume to 1L, stirs evenly, and measures pH value, the electrical conductivity of solution, up to cleaning solution after filtering.
The present invention have developed a whole set of and step auspicious BC-5800 cellanalyzers replacement reagent.
The dilution is that blood cell analysis with dilution, the LEOI hemolytic agents is blood cell analysis hemolytic agent, institute
It is blood cell analysis hemolytic agent, the LBA molten that to state LEOII dyeing liquors, which be blood cell analysis with dyeing liquor, the LH hemolytic agents,
Blood agent is that blood cell analysis with hemolytic agent, the cleaning solution is blood cell analysis cleaning solution.
Current advanced five sorting technique of haemocyte, it is thin to employ Coulter principle, colorimetric method and semiconductor laser streaming
Born of the same parents' technology, can detect red blood cell, hemoglobin, blood platelet, leucocyte five classify, the parameter such as mean corpuscular volume (MCV), pass through
The number of detection human body haemocyte judges various diseases, monitors physical condition in time, reduces disease incidence probability or control
The state of an illness processed.
Need to be used together with matched reagent during the classification Blood cell analyzer detection blood cell numbers of BC-5800 five, it is supporting
Reagent major class includes dilution, hemolytic agent, dyeing liquor, cleaning solution.The main function of dilution is to provide one and blood plasma phase
The environment of same osmotic pressure, electrical conductivity, LEOI hemolytic agents are lysed erythrocytes, keep leucocyte integrality, leucocyte is divided
Class is with counting, and LEOII dyeing liquors carry out chemical staining to cell, to observe；LH hemolytic agents are lysed erythrocytes, discharge bleeding
Lactoferrin, forms stable haemoglobin dervative with chelating agent, carries out the measure of hemoglobin concentration；LBA hemolytic agents are single
An only passage surveys basophilic granulocyte number；Cleaning solution, main function are Cleaning pipes, are mutually polluted between sample.
Beneficial effects of the present invention：
(1) product of the invention can step the original-pack reagents of auspicious BC-5800 as the classification cytoanalyze of the prior art five
Replacement reagent, can efficiently solve the problem of of high cost, delivery cycle is long；
(2) matched reagent accuracy of the invention it is high, it is reproducible, reached with genuine reagent linear regression coefficient R
More than 0.9900, the preferable correlation of system；
(3) raw material sources of the invention are extensive, are easy to get, and cheap, nontoxic, environmental pollution is few；
(4) simple production process of product of the present invention, is easy to duplication of production, and stability is good, long shelf-life, is worth further
It is widely popularized.
Brief description of the drawings
Fig. 1 is that the test result for the WBC projects that reagent made from the embodiment of the present invention 1 is surveyed respectively with stepping auspicious reagent contrasts
Correlation linear graph；
Fig. 2 is that the test result for the RBC projects that reagent made from the embodiment of the present invention 1 is surveyed respectively with stepping auspicious reagent contrasts
Correlation linear graph；
Fig. 3 is that the test result for the HGB projects that reagent made from the embodiment of the present invention 1 is surveyed respectively with stepping auspicious reagent contrasts
Correlation linear graph；
Fig. 4 is that the test result for the MCV projects that reagent made from the embodiment of the present invention 1 is surveyed respectively with stepping auspicious reagent contrasts
Correlation linear graph；
Fig. 5 is that the test result for the PLT projects that reagent made from the embodiment of the present invention 1 is surveyed respectively with stepping auspicious reagent contrasts
Correlation linear graph.
Embodiment
In order to preferably explain the present invention, it is described further in conjunction with specific examples below, but the present invention is unlimited
In specific embodiment.
Instrument：Auspicious BC-5800 cellanalyzers advanced in years, 1；
Step auspicious genuine reagent：M-58D dilutions, M-58LEO (I) hemolytic agent, M-58LEO (II) hemolytic agent, M-58LBA are molten
Blood agent, M-58LH hemolytic agents, M-58 cleaning solutions；
Reagent of the present invention：Dilution, LEOI hemolytic agents, LEOII dyeing liquors, LBA hemolytic agents, LH hemolytic agents, cleaning solution.
Embodiment 1
BC-5800 cellanalyzers replacement reagent of the present invention
(1) formula of dilution of the present invention：
Required material is accurately weighed by above-mentioned formula ratio, the imidazoles weighed, sweet ammonia are sequentially added in 1000mL beakers
Acid, sodium chloride, sodium sulphate, EDTA-2Na, add suitable deionized water, and heating stirring is completely dissolved it, and material is completely molten
Xie Hou, is cooled to less than 40 DEG C, adds the preservative weighed up, stirs evenly, the volume of constant volume to 1L, measure the pH value of solution
For 6.91, electrical conductivity 16.70ms/cm, osmotic pressure 282mOsm/kg, up to dilution after filtering.
(2) formula of LEOI hemolytic agents of the present invention：
Required material is accurately weighed by above-mentioned formula ratio, the ten alkyl front threes weighed are sequentially added in 1000mL beakers
Ammonium chloride, OP-10, PEG4000, Trisbase, P-hydroxybenzoic acid, sodium chloride, sodium sulphate, add suitable deionization
Water, heating stirring are completely dissolved it, and methyl green is added after cooling, add remaining deionized water, the volume of constant volume to 1L, is stirred
Mix uniformly, the pH value for measuring solution is 5.55, electrical conductivity 9.56ms/cm, osmotic pressure 215mOsm/kg, after filtering to obtain the final product
LEOI hemolytic agents.
(3) formula of LEOII dyeing liquors of the present invention：
Required material is accurately weighed by above-mentioned formula ratio, the dodecyl three weighed is sequentially added in 1000mL beakers
Ammonio methacrylate, TX-100, P-hydroxybenzoic acid, hydrogen phthalate sodium, sodium chloride, add suitable deionized water, heating
Stirring is completely dissolved it, adds remaining deionized water, the volume of constant volume to 1L, stirs evenly, and the pH value for measuring solution is
5.60, electrical conductivity 6.5ms/cm, osmotic pressure 130mOsm/kg, up to LEOII dyeing liquors after filtering.
(4) formula of LH hemolytic agents of the present invention：
Required material is accurately weighed by above-mentioned formula ratio, the ten alkyl front threes weighed are sequentially added in 1000mL beakers
Ammonium chloride, Brij35, sodium chloride, add suitable deionized water, and heating stirring is completely dissolved it, add and weigh up after cooling
Formic acid, stirring, add remaining deionized water, the volume of constant volume to 1L, stirs evenly, measure solution pH value be 2.17,
Electrical conductivity is 36.31ms/cm, osmotic pressure 750mOsm/kg, up to LH hemolytic agents after filtering.
(5) formula of LBA hemolytic agents of the present invention：
Required material is accurately weighed by above-mentioned formula ratio, the dodecyl three weighed is sequentially added in 1000mL beakers
Ammonio methacrylate, barbiturates, Trisbase, sodium chloride, add suitable deionized water, and heating stirring is completely dissolved it,
After cooling, chlorophenol red dissolving is added, adds remaining deionized water, the volume of constant volume to 1L, stirs evenly, and measures the pH of solution
It is worth for 9.56, electrical conductivity 14.31ms/cm, osmotic pressure 456mOsm/kg, up to LBA hemolytic agents after filtering.
(6) formula of cleaning solution of the present invention：
Accurately weigh required material by above-mentioned formula ratio, sequentially added in 1000mL beakers weighed PE6800,
AEO-9, imidazoles, sodium chloride, sodium sulphate, add suitable deionized water, and heating stirring is completely dissolved it, are added after cooling folded
Sodium nitride dissolves, and adds remaining deionized water, and the volume of constant volume to 1L, stirs evenly, and the pH value for measuring solution is 9.10, electricity
Conductance is 16.5ms/cm, up to cleaning solution after filtering.
Experimental method and result：
By BC-5800 cellanalyzers replacement reagent made from the embodiment of the present invention 1 with stepping auspicious original-pack BC M58 systems
Row reagent for hemocyte analyzers according to NCCLS files EP9-A2, (ratify by American National Clinical Laboratory Standard committee paper
Guide-the second edition-carry out method with patient's sample compares bias evaluation) method is fresh to same 101 (monstrosities containing WBC)
Blood specimen is tested, and obtains 101 groups of data, and data are listed in Table 1 below, and is tried with stepping auspicious original-pack BC M58 series cellanalyzer
Agent test result is abscissa, using BC-5800 cellanalyzers replacement reagent test result made from the embodiment of the present invention 1 as
Ordinate, linear regression is done by Origin softwares, obtains reagent made from the embodiment of the present invention 1 of Fig. 1 with stepping auspicious reagent point
The test result contrast correlation linear graph of other surveyed WBC projects, linear regression is drawn by the WBC project testing data of table 1：
Y=1.00696X-0.04163, R=0.99775；
By BC-5800 cellanalyzers replacement reagent made from the embodiment of the present invention 1 with stepping auspicious original-pack BC M58 systems
Row reagent for hemocyte analyzers according to NCCLS files EP9-A2, (ratify by American National Clinical Laboratory Standard committee paper
Guide-the second edition-carry out method with patient's sample compares bias evaluation) method is fresh to same 101 (monstrosities containing RBC)
Blood specimen is tested, and obtains 101 groups of data, and data are listed in Table 1 below, and is tried with stepping auspicious original-pack BC M58 series cellanalyzer
Agent test result is abscissa, using BC-5800 cellanalyzers replacement reagent test result made from the embodiment of the present invention 1 as
Ordinate, linear regression is done by Origin softwares, obtains reagent made from the embodiment of the present invention 1 of Fig. 2 with stepping auspicious reagent point
The test result contrast correlation linear graph of other surveyed RBC projects, linear regression is drawn by the RBC project testing data of table 1：
Y=0.99345X-0.0222, R=0.995；
By BC-5800 cellanalyzers replacement reagent made from the embodiment of the present invention 1 with stepping auspicious original-pack BC M58 systems
Row reagent for hemocyte analyzers according to NCCLS files EP9-A2, (ratify by American National Clinical Laboratory Standard committee paper
Guide-the second edition-carry out method with patient's sample compares bias evaluation) method is fresh to same 101 (monstrosities containing HGB)
Blood specimen is tested, and obtains 101 groups of data, and data are listed in Table 1 below, and is tried with stepping auspicious original-pack BC M58 series cellanalyzer
Agent test result is abscissa, using BC-5800 cellanalyzers replacement reagent test result made from the embodiment of the present invention 1 as
Ordinate, linear regression is done by Origin softwares, obtains reagent made from the embodiment of the present invention 1 of Fig. 3 with stepping auspicious reagent point
The test result contrast correlation linear graph of other surveyed HGB projects, linear regression is drawn by the HGB project testing data of table 1：
Y=1.0015X-0.52003, R=0.99486；
By BC-5800 cellanalyzers replacement reagent made from the embodiment of the present invention 1 with stepping auspicious original-pack BC M58 systems
Row reagent for hemocyte analyzers according to NCCLS files EP9-A2, (ratify by American National Clinical Laboratory Standard committee paper
Guide-the second edition-carry out method with patient's sample compares bias evaluation) method is fresh to same 101 (monstrosities containing MCV)
Blood specimen is tested, and obtains 101 groups of data, and data are listed in Table 1 below, and is tried with stepping auspicious original-pack BC M58 series cellanalyzer
Agent test result is abscissa, using BC-5800 cellanalyzers replacement reagent test result made from the embodiment of the present invention 1 as
Ordinate, linear regression is done by Origin softwares, obtains reagent made from the embodiment of the present invention 1 of Fig. 4 with stepping auspicious reagent point
The test result contrast correlation linear graph of other surveyed MCV projects, linear regression is drawn by the MCV project testing data of table 1：
Y=0.97902X+2.54324, R=0.97414；
By BC-5800 cellanalyzers replacement reagent made from the embodiment of the present invention 1 with stepping auspicious original-pack BC M58 systems
Row reagent for hemocyte analyzers according to NCCLS files EP9-A2, (ratify by American National Clinical Laboratory Standard committee paper
Guide-the second edition-carry out method with patient's sample compares bias evaluation) method is fresh to same 101 (monstrosities containing PLT)
Blood specimen is tested, and obtains 101 groups of data, and data are listed in Table 1 below, and is tried with stepping auspicious original-pack BC M58 series cellanalyzer
Agent test result is abscissa, using BC-5800 cellanalyzers replacement reagent test result made from the embodiment of the present invention 1 as
Ordinate, linear regression is done by Origin softwares, obtains reagent made from the embodiment of the present invention 1 of Fig. 5 with stepping auspicious reagent point
The test result contrast correlation linear graph of other surveyed PLT projects, linear regression is drawn by the PLT project testing data of table 1：
Y=0.98842X+3.72021, R=0.99252；
Table 1：BC-5800 cellanalyzers replacement reagent made from the embodiment of the present invention 1 is with stepping auspicious original-pack BC M58
(wherein (i) is the experiment raw data results for WBC, RBC, HGB, MCV, PLT project that serial cellanalyzer reagent place surveys
The data of auspicious original-pack BC M-58 series of cell analyzer reagents are stepped, (I) is BC-5800 haemocytes made from the embodiment of the present invention 1
The data of analyzer replacement reagent).
The foregoing is merely the specific embodiment of the present invention, it is not intended to limit the scope of the invention, every utilization
The equivalent transformation that the present invention makees, is directly or indirectly used in other relevant technical fields, is similarly included in the present invention's
Among scope of patent protection.
Claims (10)
- A kind of 1. reagent for hemocyte analyzers, it is characterised in that it include dilution, LEOI hemolytic agents, LEOII dyeing liquors, LH hemolytic agents, LBA hemolytic agents, cleaning solution；The dilution includes 2.5~8.0g/L of sodium chloride, 3.5~8.0g/L of sodium sulphate, 0.5~5.0g/L of imidazoles, glycine 0.1~3.0g/L, 0.3~1.0g/L of EDTA-2Na, 0.3~1.0g/L of preservative；The LEOI hemolytic agents include 1.0~6.0g/L of decyl trimethyl ammonium chloride, 1.0~4.0g/L of OP-10, PEG4000 1.0~6.0g/L, 2.0~10.0g/L of P-hydroxybenzoic acid, 1.0~10.0g/L of Trisbase, 2.0~10.0g/ of sodium chloride L, 2~5.0g/L of sodium sulphate, 1.0~3.0g/L of methyl green；The LEOII dyeing liquors include 1.0~4.0g/L of dodecyl trimethyl ammonium chloride, 0.5~4.0g/L of TX-100, right 0.5~5.0g/L of hydroxybenzoic acid, 2.0~5.0g/L of hydrogen phthalate sodium, 1.0~5.0g/L of sodium chloride；The LH hemolytic agents include 5.0~20.0g/L of decyl trimethyl ammonium chloride, 1.0~5.0g/L of Brij35, formic acid 1.0 ~8.0g/L, 10.0~25.0g/L of sodium chloride；The LBA hemolytic agents include 2.0~10.0g/L of dodecyl trimethyl ammonium chloride, 0.5~3.0g/L of barbiturates, 1.0~10.0g/L of Trisbase, 10.0~20.0g/L of sodium chloride, 5~10mg/L of chlorophenol red；The cleaning solution include 0.5~10.0g/L of PE6800,0.5~10.0g/L of AEO-9,1.0~10.0g/L of sodium chloride, 1.0~10.0g/L of sodium sulphate, 0.5~5.0g/L of imidazoles, 0.1~1.0g/L of sodium azide.
- 2. reagent for hemocyte analyzers according to claim 1, it is characterised in that the pH value of the dilution is 6.5 ~7.0, the electrical conductivity of the dilution is 14.0~17.0ms/cm, and the osmotic pressure of the dilution is 260~320mOsm/ kg。
- 3. reagent for hemocyte analyzers according to claim 1 or 2, it is characterised in that title is sequentially added in beaker Imidazoles, glycine, sodium chloride, sodium sulphate, the EDTA-2Na taken, add deionized water, heating stirring after it is completely dissolved, Less than 40 DEG C are cooled to, the preservative weighed up is added, stirs evenly, with the volume of deionized water constant volume to 1L, is after filtering Obtain the dilution.
- 4. reagent for hemocyte analyzers according to claim 1, it is characterised in that the pH value of the LEOI hemolytic agents is 5.50~6.0, the electrical conductivity of the LEOI hemolytic agents is 9.0~10.0ms/cm, and the osmotic pressure of the LEOI hemolytic agents is 200 ~250mOsm/kg.
- 5. the reagent for hemocyte analyzers according to claim 1 or 4, it is characterised in that title is sequentially added in beaker Decyl trimethyl ammonium chloride, OP-10, PEG4000, Trisbase, P-hydroxybenzoic acid, sodium chloride, the sodium sulphate taken, adds Enter deionized water, heating stirring adds methyl green after it is completely dissolved, equal with the volume of deionized water constant volume to 1L, stirring It is even, up to the LEOI hemolytic agents after filtering.
- 6. reagent for hemocyte analyzers according to claim 1, it is characterised in that the pH value of the LEOII dyeing liquors For 5.50~6.0, the electrical conductivity of the LEOII dyeing liquors is 6.0~7.0ms/cm, and the osmotic pressure of the LEOII dyeing liquors is 100~150mOsm/kg.
- 7. the reagent for hemocyte analyzers according to claim 1 or 6, it is characterised in that title is sequentially added in beaker The dodecyl trimethyl ammonium chloride that has taken, TX-100, P-hydroxybenzoic acid, hydrogen phthalate sodium, sodium chloride, sodium sulphate, Deionized water is added, heating stirring is completely dissolved it, with the volume of deionized water constant volume to 1L, stirs evenly, after filtering i.e. Obtain the LEOII dyeing liquors.
- 8. reagent for hemocyte analyzers according to claim 1, it is characterised in that the pH value of the LH hemolytic agents is 2.0~3.0, the electrical conductivity of the LH hemolytic agents is 35.0~40.0ms/cm, the osmotic pressure of the LH hemolytic agents for 700~ 800mOsm/kg。
- 9. reagent for hemocyte analyzers according to claim 1, it is characterised in that the pH value of the LBA hemolytic agents is 9.0~11.0, the electrical conductivity of the LBA hemolytic agents is 13.0~16.0ms/cm, the osmotic pressure of the LBA hemolytic agents for 400~ 500mOsm/kg。
- 10. reagent for hemocyte analyzers according to claim 1, it is characterised in that the pH value of the cleaning solution is 8.0 ~10.0, the electrical conductivity of the cleaning solution is 15.0~20.0ms/cm.
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| CN201711170124.4A CN107991151A (en) | 2017-11-22 | 2017-11-22 | A kind of reagent for hemocyte analyzers |
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| CN201711170124.4A CN107991151A (en) | 2017-11-22 | 2017-11-22 | A kind of reagent for hemocyte analyzers |
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Cited By (3)
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| CN109238801A (en) * | 2018-10-19 | 2019-01-18 | 武汉百合龙腾生物科技有限责任公司 | A kind of blood cell analysis hemolytic agent |
| CN109655604A (en) * | 2018-12-27 | 2019-04-19 | 山东博科生物产业有限公司 | A set of universal three classification blood cell analysis reagent |
| CN111521834A (en) * | 2020-03-20 | 2020-08-11 | 佛山市顺德区德维医疗器械有限公司 | Reagent for XN series full-automatic modular blood body fluid analyzer |
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| CN109238801A (en) * | 2018-10-19 | 2019-01-18 | 武汉百合龙腾生物科技有限责任公司 | A kind of blood cell analysis hemolytic agent |
| CN109238801B (en) * | 2018-10-19 | 2021-08-13 | 武汉百合龙腾生物科技有限责任公司 | Hemolytic agent for blood cell analysis |
| CN109655604A (en) * | 2018-12-27 | 2019-04-19 | 山东博科生物产业有限公司 | A set of universal three classification blood cell analysis reagent |
| CN111521834A (en) * | 2020-03-20 | 2020-08-11 | 佛山市顺德区德维医疗器械有限公司 | Reagent for XN series full-automatic modular blood body fluid analyzer |
| CN111521834B (en) * | 2020-03-20 | 2023-03-24 | 佛山市顺德区德维医疗科技有限公司 | Reagent for XN series full-automatic modular blood body fluid analyzer |
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