CN103290097A - Gamma-glutamoyl transferase detection reagent - Google Patents
Gamma-glutamoyl transferase detection reagent Download PDFInfo
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- CN103290097A CN103290097A CN2013102004731A CN201310200473A CN103290097A CN 103290097 A CN103290097 A CN 103290097A CN 2013102004731 A CN2013102004731 A CN 2013102004731A CN 201310200473 A CN201310200473 A CN 201310200473A CN 103290097 A CN103290097 A CN 103290097A
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- damping fluid
- gamma
- detection reagent
- glutamyl
- acid
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- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 56
- 238000001514 detection method Methods 0.000 title claims abstract description 31
- 102000004357 Transferases Human genes 0.000 title claims abstract description 16
- 108090000992 Transferases Proteins 0.000 title claims abstract description 16
- 238000006243 chemical reaction Methods 0.000 claims abstract description 24
- 239000003085 diluting agent Substances 0.000 claims abstract description 19
- 239000007788 liquid Substances 0.000 claims abstract description 9
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Natural products OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims abstract description 6
- 108090000854 Oxidoreductases Proteins 0.000 claims abstract description 6
- 102000004316 Oxidoreductases Human genes 0.000 claims abstract description 6
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 6
- 230000001681 protective effect Effects 0.000 claims abstract description 5
- 239000012530 fluid Substances 0.000 claims description 48
- 238000013016 damping Methods 0.000 claims description 44
- 125000002642 gamma-glutamyl group Chemical group 0.000 claims description 16
- 239000000463 material Substances 0.000 claims description 14
- BPYKTIZUTYGOLE-IFADSCNNSA-N Bilirubin Chemical compound N1C(=O)C(C)=C(C=C)\C1=C\C1=C(C)C(CCC(O)=O)=C(CC2=C(C(C)=C(\C=C/3C(=C(C=C)C(=O)N\3)C)N2)CCC(O)=O)N1 BPYKTIZUTYGOLE-IFADSCNNSA-N 0.000 claims description 10
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 10
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 6
- ORTQZVOHEJQUHG-UHFFFAOYSA-L copper(II) chloride Chemical compound Cl[Cu]Cl ORTQZVOHEJQUHG-UHFFFAOYSA-L 0.000 claims description 6
- SUMDYPCJJOFFON-UHFFFAOYSA-N isethionic acid Chemical compound OCCS(O)(=O)=O SUMDYPCJJOFFON-UHFFFAOYSA-N 0.000 claims description 6
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical class [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 claims description 6
- IHPYMWDTONKSCO-UHFFFAOYSA-N 2,2'-piperazine-1,4-diylbisethanesulfonic acid Chemical compound OS(=O)(=O)CCN1CCN(CCS(O)(=O)=O)CC1 IHPYMWDTONKSCO-UHFFFAOYSA-N 0.000 claims description 5
- 235000010323 ascorbic acid Nutrition 0.000 claims description 5
- 239000011668 ascorbic acid Substances 0.000 claims description 5
- 229960005070 ascorbic acid Drugs 0.000 claims description 5
- 229930182470 glycoside Natural products 0.000 claims description 5
- 150000002338 glycosides Chemical class 0.000 claims description 5
- 230000002452 interceptive effect Effects 0.000 claims description 5
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical class COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 claims description 5
- 229920000136 polysorbate Polymers 0.000 claims description 5
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 5
- 239000000758 substrate Substances 0.000 claims description 5
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 4
- 239000013543 active substance Substances 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 claims description 4
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 claims description 4
- 229960005486 vaccine Drugs 0.000 claims description 4
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 3
- CHHHXKFHOYLYRE-UHFFFAOYSA-M 2,4-Hexadienoic acid, potassium salt (1:1), (2E,4E)- Chemical compound [K+].CC=CC=CC([O-])=O CHHHXKFHOYLYRE-UHFFFAOYSA-M 0.000 claims description 3
- 108010015428 Bilirubin oxidase Proteins 0.000 claims description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 3
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 3
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 3
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims description 3
- 239000004327 boric acid Substances 0.000 claims description 3
- 229940098773 bovine serum albumin Drugs 0.000 claims description 3
- 229960003280 cupric chloride Drugs 0.000 claims description 3
- YAGKRVSRTSUGEY-UHFFFAOYSA-N ferricyanide Chemical compound [Fe+3].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] YAGKRVSRTSUGEY-UHFFFAOYSA-N 0.000 claims description 3
- 239000004292 methyl p-hydroxybenzoate Substances 0.000 claims description 3
- 235000010270 methyl p-hydroxybenzoate Nutrition 0.000 claims description 3
- 229940051841 polyoxyethylene ether Drugs 0.000 claims description 3
- 229920000056 polyoxyethylene ether Polymers 0.000 claims description 3
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 3
- 229920000053 polysorbate 80 Polymers 0.000 claims description 3
- 229910052700 potassium Inorganic materials 0.000 claims description 3
- 239000011591 potassium Substances 0.000 claims description 3
- 239000004302 potassium sorbate Substances 0.000 claims description 3
- 235000010241 potassium sorbate Nutrition 0.000 claims description 3
- 229940069338 potassium sorbate Drugs 0.000 claims description 3
- WXMKPNITSTVMEF-UHFFFAOYSA-M sodium benzoate Chemical compound [Na+].[O-]C(=O)C1=CC=CC=C1 WXMKPNITSTVMEF-UHFFFAOYSA-M 0.000 claims description 3
- 239000004299 sodium benzoate Substances 0.000 claims description 3
- 235000010234 sodium benzoate Nutrition 0.000 claims description 3
- XOGGUFAVLNCTRS-UHFFFAOYSA-N tetrapotassium;iron(2+);hexacyanide Chemical compound [K+].[K+].[K+].[K+].[Fe+2].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] XOGGUFAVLNCTRS-UHFFFAOYSA-N 0.000 claims description 3
- -1 (2-hydroxyethyl) amino Chemical group 0.000 claims description 2
- CKQPAVCUJDZLLE-UHFFFAOYSA-N 1,3-dihydroxy-2-(methylamino)butane-2-sulfonic acid Chemical compound CNC(CO)(C(C)O)S(O)(=O)=O CKQPAVCUJDZLLE-UHFFFAOYSA-N 0.000 claims description 2
- UPMBDJWMINKVIJ-UHFFFAOYSA-N 2-amino-3-hydroxy-2-methylpropane-1-sulfonic acid Chemical compound OCC(N)(C)CS(O)(=O)=O UPMBDJWMINKVIJ-UHFFFAOYSA-N 0.000 claims description 2
- XMGKYXFDYYTZFD-UHFFFAOYSA-N 4-amino-1-hydroxy-2-methylbutane-2-sulfonic acid Chemical compound OCC(C)(S(O)(=O)=O)CCN XMGKYXFDYYTZFD-UHFFFAOYSA-N 0.000 claims description 2
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 claims description 2
- QFOHBWFCKVYLES-UHFFFAOYSA-N Butylparaben Chemical compound CCCCOC(=O)C1=CC=C(O)C=C1 QFOHBWFCKVYLES-UHFFFAOYSA-N 0.000 claims description 2
- OWXMKDGYPWMGEB-UHFFFAOYSA-N HEPPS Chemical compound OCCN1CCN(CCCS(O)(=O)=O)CC1 OWXMKDGYPWMGEB-UHFFFAOYSA-N 0.000 claims 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 claims description 2
- XPJVKCRENWUEJH-UHFFFAOYSA-N Isobutylparaben Chemical compound CC(C)COC(=O)C1=CC=C(O)C=C1 XPJVKCRENWUEJH-UHFFFAOYSA-N 0.000 claims description 2
- CMHMMKSPYOOVGI-UHFFFAOYSA-N Isopropylparaben Chemical compound CC(C)OC(=O)C1=CC=C(O)C=C1 CMHMMKSPYOOVGI-UHFFFAOYSA-N 0.000 claims description 2
- 229930006000 Sucrose Natural products 0.000 claims description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 2
- 239000002253 acid Substances 0.000 claims description 2
- 229910000365 copper sulfate Inorganic materials 0.000 claims description 2
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 2
- QDPMLKBAQOZXEF-UHFFFAOYSA-N ethanesulfonic acid;sodium Chemical compound [Na].CCS(O)(=O)=O QDPMLKBAQOZXEF-UHFFFAOYSA-N 0.000 claims description 2
- 239000004403 ethyl p-hydroxybenzoate Substances 0.000 claims description 2
- 235000010228 ethyl p-hydroxybenzoate Nutrition 0.000 claims description 2
- 229940043351 ethyl-p-hydroxybenzoate Drugs 0.000 claims description 2
- NUVBSKCKDOMJSU-UHFFFAOYSA-N ethylparaben Chemical compound CCOC(=O)C1=CC=C(O)C=C1 NUVBSKCKDOMJSU-UHFFFAOYSA-N 0.000 claims description 2
- 235000011187 glycerol Nutrition 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 235000010232 propyl p-hydroxybenzoate Nutrition 0.000 claims description 2
- 239000004405 propyl p-hydroxybenzoate Substances 0.000 claims description 2
- 229960003415 propylparaben Drugs 0.000 claims description 2
- 229910052708 sodium Inorganic materials 0.000 claims description 2
- 239000011734 sodium Substances 0.000 claims description 2
- 235000010288 sodium nitrite Nutrition 0.000 claims description 2
- CIJQGPVMMRXSQW-UHFFFAOYSA-M sodium;2-aminoacetic acid;hydroxide Chemical compound O.[Na+].NCC([O-])=O CIJQGPVMMRXSQW-UHFFFAOYSA-M 0.000 claims description 2
- 239000005720 sucrose Substances 0.000 claims description 2
- 102000004190 Enzymes Human genes 0.000 claims 3
- 108090000790 Enzymes Proteins 0.000 claims 3
- 230000035945 sensitivity Effects 0.000 abstract description 7
- 238000004108 freeze drying Methods 0.000 abstract description 2
- 235000019154 vitamin C Nutrition 0.000 abstract description 2
- 239000011718 vitamin C Substances 0.000 abstract description 2
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- 230000002335 preservative effect Effects 0.000 abstract 2
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 abstract 1
- 229930003268 Vitamin C Natural products 0.000 abstract 1
- 239000000654 additive Substances 0.000 abstract 1
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- 239000004094 surface-active agent Substances 0.000 abstract 1
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- 238000010790 dilution Methods 0.000 description 4
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- YMAWOPBAYDPSLA-UHFFFAOYSA-N glycylglycine Chemical compound [NH3+]CC(=O)NCC([O-])=O YMAWOPBAYDPSLA-UHFFFAOYSA-N 0.000 description 4
- KZZWQCKYLNIOBT-UHFFFAOYSA-N 5-amino-2-nitrobenzoic acid Chemical compound NC1=CC=C([N+]([O-])=O)C(C(O)=O)=C1 KZZWQCKYLNIOBT-UHFFFAOYSA-N 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
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- IEQAICDLOKRSRL-UHFFFAOYSA-N 2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2-dodecoxyethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethanol Chemical compound CCCCCCCCCCCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCO IEQAICDLOKRSRL-UHFFFAOYSA-N 0.000 description 1
- 208000022309 Alcoholic Liver disease Diseases 0.000 description 1
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- 229920001213 Polysorbate 20 Polymers 0.000 description 1
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Abstract
The invention discloses a gamma-glutamoyl transferase detection reagent which comprises a diluent and a reaction reagent, wherein the diluent is a buffer liquid, a surfactant, a preservative, a debilirubin and a vitamin C oxidase, and the reaction reagent is a buffer liquid, a bi-glycoside peptide, L-gamma-glutamoyl-3-hydroxyl-4-nitroaniline, a primer protector, a preservative and a freeze-drying protective additive. The detection reagent disclosed by the invention has good sensitivity, accuracy, precision and linearity, and can completely satisfy the clinical examination requirement.
Description
Technical field
The present invention relates to the medical test technical field, be specifically related to a kind of gamma-glutamyl based transferase detection reagent for the POCT analyser.
Background technology
Hepatopathy is the great disease of a kind of common hazardness; its timely diagnosis and prevention and the diagnosis and treatment for the treatment of for disease are played an important role; gamma-glutamyl based transferase (γ-glutamyltransferase; γ-GGT) be distributed widely in the tissue; maximum in the kidney; secondly be pancreas and liver; γ-GGT is mainly from liver in the normal human serum; it is in acute hepatitis; chronic active hepatitis and liver cirrhosis are lost when compensatory and are only slightly raise; but when obstructive jaundice; because the acatharsia adverse current is gone into blood, during primary hepatocarcinoma, synthetic hyperfunction in liver; all can significantly raise; even reaching normally more than 10 times, alcoholic γ-GGT also obviously raises, and helps to diagnose alcoholic liver disease.γ-GGT generally adopts various large automatic Biochemical Analyzers to detect at present, but because large automatic Biochemical Analyzer equipment price height, and complicated operation, operator need have relevant expertise and accept corresponding training, and it is high to use complementary conditions to require, and need to be equipped with voltage stabilized source, water purification machine etc., and floor space is big, the maintenance cost height needs professional's time-based maintenance, so basic medical unit or household person all do not have condition to buy and use.In addition, the large hospital patient is many, detects that formality is loaded down with trivial details, long flow path, waiting time be long, and this has also brought the huge time to bear to the patient, the more important thing is that patient is affected adversely treatment because can not in time diagnosing, even can lose one's life.
(point-of-caretesting is an emerging technical field POCT) to real-time test, and principal feature is to obtain a result fast, and is easy and simple to handle, uses easily and miniaturization.Along with diagnosis and the progress of ancillary technique, and people are to the understanding of disease and the requirement for the treatment of level raising, and POCT receives publicity gradually.In fact POCT becomes the important research project of external diagnosis reagent and instrument field gradually in American-European market, existing many products put goods on the market, but this class POCT instrument has a major defect at present, be exactly that analyser and reagent consumptive material are expensive especially, has huge primary care system like this for China, and the reagent usage quantity is country greatly, and instrument and the matched reagent consumptive material of American-European exploitation obviously all face the challenge at present.
The micro-fluidic chip technology is one of of paramount importance cutting edge technology in the 21 century world, it is integrated into basic operation units such as specimen preparation related in the fields such as biological and chemical, reaction, separation, detection and cell cultures, sorting, cracking on the chip of one tens square centimeters (even littler), form network by the microchannel, running through total system with controlled fluid, is a kind of technology in order to the various functions that replace conventional biological or chemical laboratory.The micro-fluidic chip technology is incorporated into POCT equipment, started the new situation of POCT development, can make the complicated whole blood in laboratory in the past quantitatively, blood cell serum separates, serum dilution and step such as mensuration synchronously, finishes in the on-line automatic equalization of chip, reaches multiple mark synchronous detection purpose.POCT analyser in conjunction with the micro-fluidic chip technology has split hair caccuracy, low blood volume, simple to operate, few, the low cost and other advantages of detection reagent consumption of needing concurrently, as protect and give birth to the international POCT instrument of curing company limited of giving birth to, the Piccolo of Abaxis company, the Afinion of Axis-Shield company etc., this type of POCT analyser can realize that all but small amount of sample can be analyzed, easy and simple to handle, no crossed contamination automated job, is highly suitable for China and has huge primary care system and reagent usage quantity country greatly like this.
Along with Eleventh Five-Year Plan plan purchasing and strengthen at three grades and second-grade hospital infrastructure device, middle rank possesses good software and hardware facilities to go to the hospital at present, but basic medical unit particularly its full-automatic biochemical testing instruments facility of unit such as commune hospital, clinic is generally not enough, the check professional who does not also have enough numbers, therefore setting up operation POCT analytical system simple and easy, cheap, real-time report has important meaning to bringing into play the effect of vast basic hospital in disease prevention and diagnosis and treatment.And provide a kind of determination of amylase reagent that can be used for the POCT analyser of above-mentioned introducing micro-fluidic chip technology to become problem demanding prompt solution.
Summary of the invention
The present invention is directed to the above-mentioned deficiency of prior art, a kind of gamma-glutamyl based transferase detection reagent that can be used for introducing the POCT analyser of micro-fluidic chip technology is provided.
The gamma-glutamyl based transferase detection reagent that the present invention can be used for above-mentioned POCT analyser (introducing the micro-fluidic chip technology) realizes by following technical scheme.
A kind of gamma-glutamyl based transferase detection reagent that can be used for the POCT analyser comprises diluent and reaction reagent, and wherein diluent is to be grouped into by following one-tenth:
Damping fluid (pH6.5-7.5) 0.01-1.0mol/L,
Tensio-active agent 0.1-10.0%(mass percent),
Sanitas 0.1-10.0%(mass percent),
Remove bilirubin agent interfering 1-10mol/L or 1-100KU/L,
Ascorbic acid oxidase 1-100KU/L;
Wherein said reaction reagent is to be grouped into by following one-tenth:
Damping fluid (pH7.0-8.0) 0.1-1.0mol/L,
Two glycosides peptide 0.1-1mol/L,
L-γ-Gu Anxianji-3-hydroxyl-4-N-methyl-p-nitroaniline 10-100mmol/L,
Substrate protective material 1-100mmol/L,
Sanitas 0.1-1g/L,
Lyophilized vaccine 10-30g/L.
Wherein said damping fluid can be the amino damping fluid of trishydroxymethyl, glycine-NaOH damping fluid, N-2-hydroxyethyl piperazine-N'-2-ethyl sulfonic acid damping fluid, N-three (methylol) methylamino--2-hydroxy-propanesulfonic acid damping fluid, N-three (methylol) methyl-2-aminoethyl sulfonic acid damping fluid, piperazine-N, two (2-hydroxyethanesulfonic acid) damping fluids of N-, 3-morpholine-2-hydroxypropionate sodium damping fluid, 3-(N-morpholine) ethyl sulfonic acid sodium damping fluid, the 4-(2-hydroxyethyl) piperazine-1-2-hydroxy-propanesulfonic acid damping fluid, N-(2-hydroxyethyl) piperazine-N'-4-fourth sulfonic acid damping fluid, two (2-hydroxyethyl) amino of 3--2-hydroxy-propanesulfonic acid damping fluid, 3-(encircling amine)-2-hydroxyl-1-propanesulfonic acid damping fluid, 4-(2-hydroxyethyl)-1-piperazine propanesulfonic acid damping fluid, 3-(encircling amine)-1-propanesulfonic acid damping fluid, 3-morpholine propanesulfonic acid damping fluid, one or more of N-three (methylol) methyl-3-aminopropanesulfonicacid acid damping fluid.
Described tensio-active agent is nonionogenic tenside, as be selected from TWEEN series (as tween (TWEEN) 20, polysorbate40, polysorbate60, tween 80), SPAN series is (as (SPAN) 20 of class of department, class of department 40, class of department 60, class of department 80), TRITON series is (as TritonX-100, TritonX-114, TritonX-405) one or more of concrete material (namely can be TWEEN series in, a kind of concrete material in SPAN series or the TRITON series, or more than one concrete material in every kind of series, or a kind of concrete material in two kinds or the above series).
The described bilirubin agent interfering that goes is in yellow prussiate of potash, high-potassium ferricyanide, the bilirubin oxidase one or more.
Described sanitas is selected from a kind of concrete material in potassium sorbate, Sodium Benzoate, Sodium Nitrite, the proclin series sanitas (as Proclin300) or a kind of concrete material in the nipagin esters (as methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, propylparaben, butyl p-hydroxybenzoate, p-Hydroxybenzoic acid isopropyl ester, p-Hydroxybenzoic acid isobutyl ester).
Described substrate protective material is selected from one or several in dimethyl sulfoxide (DMSO), ethylene glycol, glycerine, cupric chloride, copper sulfate, the boric acid.
Described lyophilized vaccine is selected from one or several in trehalose, sucrose, bovine serum albumin, tween 80, triton x-100, the fatty alcohol-polyoxyethylene ether (Brij-35).
Diluent is liquid state in described γ-GGT mensuration reagent, and reaction reagent is dry powder.
The preparation method of diluent that described γ-GGT measures reagent is as follows: described reagent composition is added to mix behind the distilled water according to formula rate stir evenly.
The preparation method of reaction reagent that described γ-GGT measures reagent is as follows: described reagent composition is added to mix behind the distilled water according to formula rate stir evenly, 0.5-10 μ l reaction reagent is joined the reaction detection groove, volatilize 24-72h through freeze-drying (industry routine techniques) or 2-8 ℃.
The test condition that described γ-GGT measures reagent is as follows: temperature: 37 ℃; Detect predominant wavelength 405nm, commplementary wave length 750nm.
Micro-fluidic chip technology of the present invention is that basic operation unit is integrated on the chip of one tens square centimeters (even littler), forms network by the microchannel, runs through a kind of technology of total system with controlled fluid.It is two-layer up and down that its characteristics are that chip generally is divided into, the upper strata is useful on the through hole of application of sample, and lower floor comprises the respond difform fluid channel etc. of the reaction detection groove of reagent, one group of self check groove that is used for system compensation, one group of overflow groove, many group control fluid flow of sample cell, dilution liquid bath, sample quantitative slot, diluent quantitative slot, reservoir, a plurality of prepackage.Its detection method generally comprises following steps: (1) is injected into sample solution and diluent in described sample cell and the dilution liquid bath through through hole separately; (2) starter motor rotates described chip; (3) sample solution is realized solid-liquid separation and quantitative under centrifugal action, and diluent enters the diluent quantitative slot simultaneously; (4) quantified sample is mixed with diluent inflow tempering tank; (5) mixed liquid enters the reaction detection groove and reaction reagent reacts; (6) by in the reaction detection groove, carrying out in situ detection with the supporting test set of chip.
The measuring method of described γ-GGT mensuration reagent is as follows: 5-20 μ l sample joined sample cell, 20-100 μ l diluent joined the dilution liquid bath, and starter motor, record absorbance A 1 behind 37 ℃ of reaction 1min continues to record absorbance A 2 behind the reaction 5-9min.
The reaction principle that described γ-GGT measures reagent is: sample mixes with diluent earlier hatches; to remove the interfering substances such as bilirubin, vitamins C and blood fat in the sample; after mixed solution enters the reaction detection groove; be substrate with the gamma-glutamyl of the L-in the reaction reagent-3-carboxyl-p-Nitroaniline; glycylglycine is the acceptor of gamma-glutamyl; under the catalysis of γ-GGT, glutamyl is transferred on the glycylglycine molecule in sample, generates yellow 5-amino-2 nitrobenzoic acid simultaneously.And 5-amino-2 nitrobenzoic acid that generates has maximum absorption band at 405nm wavelength place, and γ-GGT's is active proportional in the speed that 5-amino-2 nitrobenzoic acid generates and the serum, therefore can be by measuring under the 405nm wavelength, the speed indicator that absorbancy increases is calculated the vigor of γ-GGT.
Advantage of the present invention and beneficial effect: reagent of the present invention has good sensitivity, accuracy, precision and linearity, can satisfy the Clinical Laboratory requirement fully.Reagent of the present invention can be used for introducing the POCT analyser of micro-fluidic chip technology, thereby realizes that operation is simple and easy, cheap, the foundation of the POCT analytical system of real-time report.
Description of drawings
Fig. 1 detects with the analyte that the high density serum specimen of collecting from hospital is diluted to different concns, the linear figure as a result of γ-GGT.
The GGT end value of measuring on Fig. 2 and the automatic clinical chemistry analyzer compares figure as a result, and wherein X-axis represents determination data in the automatic clinical chemistry analyzer Hitachi 7180, and Y-axis represents determination data on the POCT analyser.
Embodiment
To further specify the present invention by following non-limiting example below, as well known to those skilled in the art, without departing from the spirit of the invention, can make many modifications to the present invention, such modification also falls into scope of the present invention.
Following experimental technique is ordinary method if no special instructions, and employed experiment material all can easily be obtained from commercial company if no special instructions.
Embodiment 1
Diluent:
Amino damping fluid (pH7.0) 0.1mol/L of trishydroxymethyl
TWEEN?80?5%
Proclin300?0.1%
Bilirubin oxidase 1KU/L
Ascorbic acid oxidase 1KU/L
Reaction reagent:
Amino damping fluid (pH7.5) 0.15mol/L of trishydroxymethyl
Two glycosides peptide 0.1mol/L
L-γ-Gu Anxianji-3-hydroxyl-4-N-methyl-p-nitroaniline 10mmol/L
Cupric chloride 10mmol/L
Potassium sorbate 0.1g/L
Bovine serum albumin 5g/L
Embodiment 2
Diluent:
Amino damping fluid (pH7.5) 0.5mol/L of trishydroxymethyl
TRITON?X-100?5%
Methyl p-hydroxybenzoate 1%
Yellow prussiate of potash 1mol/L
Ascorbic acid oxidase 2KU/L
Reaction reagent:
3-morpholine propanesulfonic acid damping fluid (pH7.0) 0.2mol/L
Two glycosides peptide 0.2mol/L
L-γ-Gu Anxianji-3-hydroxyl-4-N-methyl-p-nitroaniline 20mmol/L
Boric acid 50 mmol/L
Proclin300?1g/L
Trehalose 10g/L
Embodiment 3
Diluent:
3-morpholine propanesulfonic acid damping fluid (pH6.5) 1.0mol/L
SPAN?20?10%
Sodium Benzoate 1%
High-potassium ferricyanide 10mol/L
Ascorbic acid oxidase 10KU/L
Reaction reagent:
Piperazine-N, two (2-hydroxyethanesulfonic acid) damping fluid (pH8.0) 1mol/L of N-
Two glycosides peptide 1mol/L
L-γ-Gu Anxianji-3-hydroxyl-4-N-methyl-p-nitroaniline 100mmol/L
DMSO?100?mmol/L
Proclin300?1g/L
Fatty alcohol-polyoxyethylene ether 10g/L
Describe below in conjunction with the performance of form to the embodiment of the invention 1 gained reagent.
1, precision
Table 1, precision assessment result
2, linearity
The analyte that is diluted to different concns with the high density serum specimen of collecting from hospital detects, and γ-GGT linearity the results are shown in Fig. 1..
3, methodology comparison test
Compare with the GGT end value of measuring on the automatic clinical chemistry analyzer, result such as Fig. 2, wherein X-axis represents determination data in the automatic clinical chemistry analyzer Hitachi 7180, POCT analyser (the living world of Taiwan guarantor of micro-fluidic chip technology is introduced in the Y-axis representative, Amishield TMO-100) goes up determination data.
4, detection sensitivity
The appraisal procedure of the detection sensitivity standard deviation of 10-20 dummy signal strength, and the standard deviation of 3-15 least significant non-zero sample signal strength are calculated gained with the EP Evaluator release6 of statistical software.Experimental result shows that reagent of the present invention has good sensitivity, can meet the improvement of U.S. clinical labororatory fully and amend legislation.
Table 2 reagent detection sensitivity
From above-mentioned detected result as can be known, reagent of the present invention has good sensitivity, accuracy, precision and linearity, can satisfy the Clinical Laboratory requirement fully.
Claims (10)
1. gamma-glutamyl based transferase detection reagent, it is characterized in that: this reagent comprises diluent and reaction reagent, wherein diluent is to be grouped into by following one-tenth:
Damping fluid (pH6.5-7.5) 0.01-1.0mol/L,
Tensio-active agent 0.1-10.0%(mass percent),
Sanitas 0.1-10.0%(mass percent),
Remove bilirubin agent interfering 1-10mol/L or 1-100KU/L,
Ascorbic acid oxidase 1-100KU/L;
Wherein said reaction reagent is to be grouped into by following one-tenth:
Damping fluid (pH7.0-8.0) 0.1-1.0mol/L,
Two glycosides peptide 0.1-1mol/L,
L-γ-Gu Anxianji-3-hydroxyl-4-N-methyl-p-nitroaniline 10-100mmol/L,
Substrate protective material 1-100mmol/L,
Sanitas 0.1-1g/L,
Lyophilized vaccine 10-30g/L.
2. gamma-glutamyl based transferase detection reagent according to claim 1; it is characterized in that: described damping fluid is the amino damping fluid of trishydroxymethyl; glycine-NaOH damping fluid; N-2-hydroxyethyl piperazine-N'-2-ethyl sulfonic acid damping fluid; N-three (methylol) methylamino--2-hydroxy-propanesulfonic acid damping fluid; N-three (methylol) methyl-2-aminoethyl sulfonic acid damping fluid; piperazine-N, two (2-hydroxyethanesulfonic acid) damping fluids of N-; 3-morpholine-2-hydroxypropionate sodium damping fluid; 3-(N-morpholine) ethyl sulfonic acid sodium damping fluid; the 4-(2-hydroxyethyl) piperazine-1-2-hydroxy-propanesulfonic acid damping fluid; N-(2-hydroxyethyl) piperazine-N'-4-fourth sulfonic acid damping fluid; two (2-hydroxyethyl) amino of 3--2-hydroxy-propanesulfonic acid damping fluid; 3-(encircling amine)-2-hydroxyl-1-propanesulfonic acid damping fluid; 4-(2-hydroxyethyl)-1-piperazine propanesulfonic acid damping fluid; 3-(encircling amine)-1-propanesulfonic acid damping fluid; 3-morpholine propanesulfonic acid damping fluid; one or more of N-three (methylol) methyl-3-aminopropanesulfonicacid acid damping fluid.
3. γ according to claim 1--glutamyl transferring enzyme detection reagent is characterized in that: described tensio-active agent is to be selected from TWEEN series, SPAN series, the TRITON series one or more of concrete material.
4. gamma-glutamyl based transferase detection reagent according to claim 1 is characterized in that: the described bilirubin agent interfering that goes is in yellow prussiate of potash, high-potassium ferricyanide, the bilirubin oxidase one or more.
5. gamma-glutamyl based transferase detection reagent according to claim 1 is characterized in that: described sanitas is a kind of in a kind of or parabens in potassium sorbate, Sodium Benzoate, Sodium Nitrite, the proclin series sanitas.
6. gamma-glutamyl based transferase detection reagent according to claim 5 is characterized in that: described proclin series sanitas is Proclin300.
7. gamma-glutamyl based transferase detection reagent according to claim 5 is characterized in that: described nipagin esters is a kind of in methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, propylparaben, butyl p-hydroxybenzoate, p-Hydroxybenzoic acid isopropyl ester, the p-Hydroxybenzoic acid isobutyl ester.
8. gamma-glutamyl based transferase detection reagent according to claim 1, it is characterized in that: described substrate protective material is one or several in dimethyl sulfoxide (DMSO), ethylene glycol, glycerine, cupric chloride, copper sulfate, the boric acid.
9. γ according to claim 1--glutamyl transferring enzyme detection reagent is characterized in that: described lyophilized vaccine is one or several in trehalose, sucrose, bovine serum albumin, tween 80, triton x-100, the fatty alcohol-polyoxyethylene ether.
10. gamma-glutamyl based transferase detection reagent according to claim 1 is characterized in that: the diluent that described γ-aminoacyl transferring enzyme is measured in the reagent is liquid state, and reaction reagent is dry powder.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104215631A (en) * | 2014-08-14 | 2014-12-17 | 苏州康铭诚业医用科技有限公司 | Buffer for determining human serum glutamyltransferase |
| CN112608977A (en) * | 2020-12-10 | 2021-04-06 | 武汉伊莱瑞特生物科技股份有限公司 | Gamma-glutamyl transpeptidase detection kit and detection method thereof |
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| CN102796804A (en) * | 2012-07-26 | 2012-11-28 | 上海迪安医学检验所有限公司 | Detection method of specific gamma-glutamyl transferase and application thereof |
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| CN102796804A (en) * | 2012-07-26 | 2012-11-28 | 上海迪安医学检验所有限公司 | Detection method of specific gamma-glutamyl transferase and application thereof |
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| 张宪等: "1118例健康人尿GGT参考值调查", 《中国医科大学学报》 * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104215631A (en) * | 2014-08-14 | 2014-12-17 | 苏州康铭诚业医用科技有限公司 | Buffer for determining human serum glutamyltransferase |
| CN112608977A (en) * | 2020-12-10 | 2021-04-06 | 武汉伊莱瑞特生物科技股份有限公司 | Gamma-glutamyl transpeptidase detection kit and detection method thereof |
| CN112608977B (en) * | 2020-12-10 | 2022-05-10 | 武汉伊莱瑞特生物科技股份有限公司 | Gamma-glutamyl transpeptidase detection kit and detection method thereof |
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