CN104198421A - Detection kit for measuring content of urea without interference of endogenous ammonia in serum - Google Patents
Detection kit for measuring content of urea without interference of endogenous ammonia in serum Download PDFInfo
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- CN104198421A CN104198421A CN201410401509.7A CN201410401509A CN104198421A CN 104198421 A CN104198421 A CN 104198421A CN 201410401509 A CN201410401509 A CN 201410401509A CN 104198421 A CN104198421 A CN 104198421A
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- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 title claims abstract description 42
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 title claims abstract description 41
- 239000004202 carbamide Substances 0.000 title claims abstract description 41
- 210000002966 serum Anatomy 0.000 title claims abstract description 38
- 238000001514 detection method Methods 0.000 title claims abstract description 36
- 229910021529 ammonia Inorganic materials 0.000 title claims abstract description 21
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 155
- BOPGDPNILDQYTO-NNYOXOHSSA-N nicotinamide-adenine dinucleotide Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 BOPGDPNILDQYTO-NNYOXOHSSA-N 0.000 claims abstract description 22
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 claims abstract description 21
- 108010046334 Urease Proteins 0.000 claims abstract description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 11
- 239000004094 surface-active agent Substances 0.000 claims abstract description 8
- 239000011780 sodium chloride Substances 0.000 claims abstract description 5
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N dimethyl sulfoxide Natural products CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 26
- KPGXRSRHYNQIFN-UHFFFAOYSA-N 2-oxoglutaric acid Chemical compound OC(=O)CCC(=O)C(O)=O KPGXRSRHYNQIFN-UHFFFAOYSA-N 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 18
- 230000002255 enzymatic effect Effects 0.000 claims description 17
- 101710088194 Dehydrogenase Proteins 0.000 claims description 16
- 238000002835 absorbance Methods 0.000 claims description 15
- 230000001681 protective effect Effects 0.000 claims description 15
- 238000013016 damping Methods 0.000 claims description 13
- 239000012530 fluid Substances 0.000 claims description 13
- 230000002421 anti-septic effect Effects 0.000 claims description 10
- HWXBTNAVRSUOJR-UHFFFAOYSA-N alpha-hydroxyglutaric acid Natural products OC(=O)C(O)CCC(O)=O HWXBTNAVRSUOJR-UHFFFAOYSA-N 0.000 claims description 9
- 229940009533 alpha-ketoglutaric acid Drugs 0.000 claims description 9
- 238000012544 monitoring process Methods 0.000 claims description 5
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 claims description 4
- 239000007993 MOPS buffer Substances 0.000 claims description 2
- 108010039918 Polylysine Proteins 0.000 claims description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 2
- 230000008859 change Effects 0.000 claims description 2
- 229920000656 polylysine Polymers 0.000 claims description 2
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 2
- 229920000053 polysorbate 80 Polymers 0.000 claims description 2
- 239000001632 sodium acetate Substances 0.000 claims description 2
- 235000017281 sodium acetate Nutrition 0.000 claims description 2
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 claims description 2
- 239000007983 Tris buffer Substances 0.000 claims 2
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 claims 1
- 239000007995 HEPES buffer Substances 0.000 claims 1
- 108090000790 Enzymes Proteins 0.000 abstract description 4
- 102000004190 Enzymes Human genes 0.000 abstract description 4
- 239000003223 protective agent Substances 0.000 abstract description 4
- KPGXRSRHYNQIFN-UHFFFAOYSA-L 2-oxoglutarate(2-) Chemical compound [O-]C(=O)CCC(=O)C([O-])=O KPGXRSRHYNQIFN-UHFFFAOYSA-L 0.000 abstract 2
- 239000007853 buffer solution Substances 0.000 abstract 2
- 239000003755 preservative agent Substances 0.000 abstract 2
- 230000002335 preservative effect Effects 0.000 abstract 2
- 101000950981 Bacillus subtilis (strain 168) Catabolic NAD-specific glutamate dehydrogenase RocG Proteins 0.000 abstract 1
- 102000016901 Glutamate dehydrogenase Human genes 0.000 abstract 1
- 238000012360 testing method Methods 0.000 description 10
- 238000002474 experimental method Methods 0.000 description 7
- 238000013459 approach Methods 0.000 description 6
- 238000003556 assay Methods 0.000 description 6
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- 238000002360 preparation method Methods 0.000 description 5
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
- QSJXEFYPDANLFS-UHFFFAOYSA-N Diacetyl Chemical group CC(=O)C(C)=O QSJXEFYPDANLFS-UHFFFAOYSA-N 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 230000007423 decrease Effects 0.000 description 4
- PCKPVGOLPKLUHR-UHFFFAOYSA-N indoxyl Chemical group C1=CC=C2C(O)=CNC2=C1 PCKPVGOLPKLUHR-UHFFFAOYSA-N 0.000 description 4
- ZWLUXSQADUDCSB-UHFFFAOYSA-N phthalaldehyde Chemical compound O=CC1=CC=CC=C1C=O ZWLUXSQADUDCSB-UHFFFAOYSA-N 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 238000006555 catalytic reaction Methods 0.000 description 3
- 239000002872 contrast media Substances 0.000 description 3
- 238000010219 correlation analysis Methods 0.000 description 3
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 3
- RHGKLRLOHDJJDR-BYPYZUCNSA-N L-citrulline Chemical compound NC(=O)NCCC[C@H]([NH3+])C([O-])=O RHGKLRLOHDJJDR-BYPYZUCNSA-N 0.000 description 2
- RHGKLRLOHDJJDR-UHFFFAOYSA-N Ndelta-carbamoyl-DL-ornithine Natural products OC(=O)C(N)CCCNC(N)=O RHGKLRLOHDJJDR-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000001569 carbon dioxide Substances 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 235000013477 citrulline Nutrition 0.000 description 2
- 229960002173 citrulline Drugs 0.000 description 2
- FSEUPUDHEBLWJY-HWKANZROSA-N diacetylmonoxime Chemical compound CC(=O)C(\C)=N\O FSEUPUDHEBLWJY-HWKANZROSA-N 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 229940054441 o-phthalaldehyde Drugs 0.000 description 2
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- 238000003908 quality control method Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- JFRMYMMIJXLMBB-UHFFFAOYSA-N xanthydrol Chemical compound C1=CC=C2C(O)C3=CC=CC=C3OC2=C1 JFRMYMMIJXLMBB-UHFFFAOYSA-N 0.000 description 2
- DYNFCHNNOHNJFG-UHFFFAOYSA-M 2-formylbenzoate Chemical compound [O-]C(=O)C1=CC=CC=C1C=O DYNFCHNNOHNJFG-UHFFFAOYSA-M 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- KRHYYFGTRYWZRS-UHFFFAOYSA-M Fluoride anion Chemical compound [F-] KRHYYFGTRYWZRS-UHFFFAOYSA-M 0.000 description 1
- 206010018364 Glomerulonephritis Diseases 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 description 1
- BAWFJGJZGIEFAR-NNYOXOHSSA-N NAD zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-N 0.000 description 1
- BAWFJGJZGIEFAR-NNYOXOHSSA-O NAD(+) Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-O 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 235000006040 Prunus persica var persica Nutrition 0.000 description 1
- 240000006413 Prunus persica var. persica Species 0.000 description 1
- 206010037601 Pyelonephritis chronic Diseases 0.000 description 1
- 208000001647 Renal Insufficiency Diseases 0.000 description 1
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 1
- MCRQDMGWGJTTJV-UHFFFAOYSA-N [C-]#N.N=O.[Na+] Chemical compound [C-]#N.N=O.[Na+] MCRQDMGWGJTTJV-UHFFFAOYSA-N 0.000 description 1
- 238000005903 acid hydrolysis reaction Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 235000009697 arginine Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003851 biochemical process Effects 0.000 description 1
- 239000012490 blank solution Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 208000020832 chronic kidney disease Diseases 0.000 description 1
- 201000006368 chronic pyelonephritis Diseases 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
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- 230000007547 defect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- -1 diazine compound Chemical class 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 208000028208 end stage renal disease Diseases 0.000 description 1
- 201000000523 end stage renal failure Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- DBLXOVFQHHSKRC-UHFFFAOYSA-N ethanesulfonic acid;2-piperazin-1-ylethanol Chemical compound CCS(O)(=O)=O.OCCN1CCNCC1 DBLXOVFQHHSKRC-UHFFFAOYSA-N 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 235000013905 glycine and its sodium salt Nutrition 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- QWPPOHNGKGFGJK-UHFFFAOYSA-N hypochlorous acid Chemical compound ClO QWPPOHNGKGFGJK-UHFFFAOYSA-N 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 201000006370 kidney failure Diseases 0.000 description 1
- 229910001425 magnesium ion Inorganic materials 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 229950006238 nadide Drugs 0.000 description 1
- KCXFHTAICRTXLI-UHFFFAOYSA-N propane-1-sulfonic acid Chemical compound CCCS(O)(=O)=O KCXFHTAICRTXLI-UHFFFAOYSA-N 0.000 description 1
- 210000002307 prostate Anatomy 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a detection kit for measuring the content of urea without the interference of endogenous ammonia in serum. The detection kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 contains 1-500mmol/L buffer solution with the pH value of 3.0-8.0, 1-80KU/L glutamate dehydrogenase, 0.5-2KU/L NADH, 1-100mmol/L alpha-oxoglutarate, 0.1-0.5% of a surface active agent and 0.1-1g/L of preservative; the reagent R2 contains 1-500mmol/L buffer solution with the pH value of 3.0-8.0, 1-60KU/L urease, 0.5-2KU/L NADH, 1-100mmol/L alpha-oxoglutarate, 10-500mmol/L sodium chloride, 1-50mmol/L enzyme protective agent, and 0.1-1g/L preservative. The kit can eliminate the interference of the endogenous ammonia and has the characteristics of excellent stability and strong anti-interference ability.
Description
Technical field
The present invention is specifically related to a kind of detection kit of the mensuration urea content that is not subject to the interference of serum endogenous ammonia.
Background technology
Urea (Urea) is for diagnosing glomerulonephritis, End Stage Renal Disease, renal failure, chronic pyelonephritis, prostate enlargement, lithangiuria, ankylurethria, tumor of bladder to cause compression of urethra etc. clinically.
The assay method of urea comprises: chemical method, urase one Podbielniak method etc.Chemical method can be divided into again: (1) Diacetylmonoxime development process, principle is: urea can with diacetyl effect, under strong acid heating condition, generate peach diazine compound, in 540nm colorimetric, unstable because of diacetyl, conventional Diacetylmonoxime replaces, and the latter meets acid hydrolysis and becomes diacetyl.This method selectivity is poor, and the range of linearity is narrow, and in reagent, contains strong chemicals, perishable instrument and contaminated environment.(2) o-phthalaldehyde(OPA) colourimetry: urea, under strong acidic environment, produces orange red product with o-phthalaldehyde(OPA) condensation, colorimetric estimation it.(3) xanthydrol turbidimetry: urea be combined with xanthydrol form infusible precipitate, colorimetric estimation it.Adopt chemical method to detect urea, specificity is all not high, disturbs as citrulline reacts just to have to diacetyl; Glycocoll, arginine, citrulline and sulfa drugs just have and disturb O-phthalic aldehyde reaction.In addition with strong acid or strong alkali environment, or need higher temperature (90 DEG C) reaction, more difficultly realize robotization, thus these class methods oneself through less use.Urase one Podbielniak method: measuring principle: first use urase hydrolyze urea, produce 2 molecules of ammonia and 1 molecule carbon dioxide.Then, ammonia reacts with phenol and hypochlorous acid in alkaline medium, generates blue indoxyl, and this process need be used nitroso-sodium cyanide catalytic reaction.The growing amount of blue indoxyl is directly proportional to urea content, in the colorimetric estimation of 630nm wavelength.The deficiency of this method is that pollution or the use ammonium salt anti-coagulants of Ammonia in Air gas to reagent or glassware can make result higher.High concentration fluoride can suppress urease, causes that result is false on the low side.Urase-glutamte dehydrogenase coupling method, by measure blank tube, standard pipe and the mensuration pipe fall off rate in 340nm place absorbance under identical conditions, can calculate the content of urea in sample.But the method is subject to the serious interference of sample endogenous ammonia, and reagent stability is poor.
Summary of the invention
The object of the invention is to overcome the defect that above-mentioned available reagent box exists, a kind of detection kit of the mensuration urea content that is not subject to the interference of serum endogenous ammonia is provided.Detect the Urea content in serum by test kit of the present invention, can reach easy and simple to handle, good stability, antijamming capability is strong, fast, measurement result object accurately and reliably.
The principle that the present invention measures Urea content is: urea is under the catalysis of urease (Urease), hydrolysis generates ammonia and carbon dioxide, ammonia is under α-ketoglutaric acid and reduced diphosphopyridine nucleotide (NADH) existence, through glutamte dehydrogenase (GLDH) catalysis, generate glutamic acid, NADH is oxidized to NAD+ simultaneously, causes the decline of 340nm wavelength place absorbance; The decline of absorbance is directly proportional to the concentration of urea in serum, by the variation of monitoring 340nm wavelength place absorbance, can measure the concentration of urea in serum.
Especially, the present invention has added glutamte dehydrogenase and NADH in R1 reagent, eliminates the interference of endogenous ammonia, to improve the antijamming capability of reagent.
Before not adding R2, endogenous ammonia elder generation and reagent R1 in sample react, and are used for eliminating the interference of endogenous ammonia, and reaction equation is:
Then after adding R2, react as follows again:
Meanwhile, basis is bright has added enzymatic protective reagent in reagent R2, has greatly improved the stability of reagent.Experiment shows, adds the reagent enzymatic activity of enzymatic protective reagent to be far superior to not add the reagent of enzyme stabilizers.
The object of the invention is to be achieved through the following technical solutions:
The detection kit that the present invention relates to a kind of mensuration urea content that is not subject to the interference of serum endogenous ammonia, comprises reagent R1 and reagent R2; Damping fluid, 1～80KU/L glutamte dehydrogenase, 0.5～2KU/L reducibility coenzyme I (NADH), 1～100mmol/L α-ketoglutaric acid, 1 ‰～5 ‰ (that is: surfactant accounts for the weight percent content of R1 gross weight) surfactant and 0.1～1g/L antiseptic that described reagent R1 contains 1～500mmol/L pH=3.0～8.0; Damping fluid, 1～60KU/L urease, 0.5～2KU/LNADH, 1～100mmol/L α-ketoglutaric acid, 10～500mmol/L sodium chloride, 1～50mmol/L enzymatic protective reagent and 0.1～1g/L antiseptic that described reagent R2 contains 1～500mmol/L pH=3.0～8.0.
Preferably, described reagent R1 contains 100～300mmol/L pH=5.0～8.0 damping fluid, 20～60KU/L glutamte dehydrogenase, 1～1.5KU/L NADH, 50～100mmol/L α-ketoglutaric acid, 1 ‰～5 ‰ surfactant and 0.2～0.8g/L antiseptic; Damping fluid, 10～30KU/L urease, 1～1.5KU/L NADH, 50～100mmol/L α-ketoglutaric acid, 100～300mmol/L sodium chloride, 10～40mmol/L enzymatic protective reagent and 0.2～0.8g/L antiseptic that described reagent R2 contains 100～300mmol/L pH=5.0～8.0.
Preferably, the damping fluid comprising in described reagent R1, R2 is selected from respectively trishydroxymethylaminomethane (TRIS), propane sulfonic acid (MOPS), 4-hydroxyethyl piperazine ethanesulfonic acid (HEPES) or BICN.
Preferred, the damping fluid comprising in described reagent R1, R2 is TRIS.Inventor's discovery, in reagent system of the present invention, this damping fluid disturbs very little to Biochemical processes, do not precipitate with calcium, magnesium ion and heavy metal ion.
Preferably, described enzymatic protective reagent is dimethyl sulfoxide (DMSO).
Preferably, described surfactant is polysorbas20, Tween 80 or triton x-100.More preferably polysorbas20.
Preferably, the antiseptic comprising in described reagent R1, R2 is selected from respectively sodium azide, proclin300, polylysine or sodium acetate.
The invention still further relates to a kind of using method of above-mentioned detection kit, reagent R1 adds after sample 37 DEG C to hatch 5 minutes, add again reagent R2, hatch 1.5 minutes for 37 DEG C, the monitoring absorbance rate of change of 3 minutes continuously, compare with urea content and the absorbance relation curve of Urea standard items, obtain the urea content of sample.
Preferably, the predominant wavelength that described monitoring adopts is 340nm, and commplementary wave length is 405nm.
Preferably, the volume ratio of described reagent R1 and reagent R2 is 4: 1.
Compared with prior art, the present invention has following beneficial effect:
1, the present invention, in order to eliminate the interference of endogenous ammonia in sample, utilizes the feature of double reagent method in automatic analysis, has added glutamte dehydrogenase and NADH in R1 reagent, and reaction can be eliminated the interference of endogenous ammonia.
2, the present invention adds enzymatic protective reagent in R2 reagent, has greatly improved reagent stability.
3, test shows, applies kit detection urea result precision of the present invention high, and precision is good.
4, kit of the present invention is applied widely, is convenient to promote the use of, and can be applicable to situation of all-level hospitals, sanitary precaution department and medical biotechnology R&D institution and measures urea in serum content.
Brief description of the drawings
By reading the detailed description of non-limiting example being done with reference to the following drawings, it is more obvious that other features, objects and advantages of the present invention will become:
Fig. 1 is the typical curve of Urea normative reference, and wherein X-axis represents the content of Urea, and Y-axis represents absorbance;
Fig. 2 is the Urea reagent that adopts respectively reagent of the present invention and Beckman company, adopt Hitachi's automatic clinical chemistry analyzer to measure by each autoregressive parameter 50 parts of human serums (comprising normal and monstrosity) simultaneously, measured value is carried out to the schematic diagram of correlation analysis; What wherein X-axis represented is patients serum's result that reagent of the present invention is measured, and what Y-axis represented is patients serum's result that Beckman company reagent is measured, coefficient R
2=0.9977, regression equation is y=1.1598x-0.7773;
Fig. 3 is reagent of the present invention and Beckman company reagent serum correlativity schematic diagram, and what wherein X-axis represented is patients serum's result that reagent of the present invention is measured, and what Y-axis represented is patients serum's result that Beckman company reagent is measured, coefficient R
2=0.9959, regression equation is y=1.0394x-0.1043;
Fig. 4 is that in kit of the present invention, R1 does not add glutamte dehydrogenase and NADH reagent and Beckman company reagent serum correlativity schematic diagram, what wherein X-axis represented is patients serum's result that R1 does not add glutamte dehydrogenase and NADH reagent mensuration, what Y-axis represented is patients serum's result that Beckman company reagent is measured, coefficient R
2=0.7881, regression equation is y=0.9147x+0.2858.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in detail.Following examples will contribute to those skilled in the art further to understand the present invention, but not limit in any form the present invention.It should be pointed out that to those skilled in the art, without departing from the inventive concept of the premise, can also make some distortion and improvement.These all belong to protection scope of the present invention.Raw material in following examples is existing conventional raw material, can directly buy acquisition by businessman.In following examples of the present invention, there is no the operation of special instruction, all can adopt existing routine techniques means.
embodiment 1
Mensuration kit is composed as follows:
1. reagent R1 is:
2. reagent R2 is:
Various compositions can add under room temperature successively, or add simultaneously, or packaging preparation immediately again before detecting separately respectively.
3. reagent detection ratio is 4: 1
The CRE detection kit that the present embodiment is described, is applicable to various types of full automatic biochemical apparatus, and taking Hitachi's 7170 full automatic biochemical apparatus as example, it operates as table 1.Analytical approach: Two point end assay, the consumption of reagent R1, R2 is respectively 240ul and 60ul, sample size 3.5ul; 240ul reagent R1 adds 3.5ul sample to hatch 5min in 37 DEG C, adds 60ul R2, hatches 1.5min for 37 DEG C, monitors continuously the variation of 180s absorbance, calculates △ A/min; Detection predominant wavelength is 340nm, and commplementary wave length is 405nm.
Adopt this reagent and said determination method, the curve (as shown in Figure 1) of the Urea standard items (C.f.a.s. of Roche company) that employing Hitachi 7170 Biochemical Analyzers record, wherein X-axis represents Urea content (mmol/L); Y-axis represents absorbance.
Table 1
embodiment 2
Mensuration kit is composed as follows:
1. reagent R1 is:
2. reagent R2 is:
Various compositions can add under room temperature successively, or add simultaneously, or packaging preparation immediately again before detecting separately respectively.
3. reagent detection ratio is 4: 1
The CRE detection kit that the present embodiment is described, is applicable to various types of full automatic biochemical apparatus, and taking Hitachi's 7170 full automatic biochemical apparatus as example, it operates as table 1.Analytical approach: Two point end assay, the consumption of reagent R1, R2 is respectively 240ul and 60ul, sample size 3.5ul; 240ul reagent R1 adds 3.5ul sample to hatch 5min in 37 DEG C, adds 60ul R2, hatches 1.5min for 37 DEG C, monitors continuously the variation of 180s absorbance, calculates △ A/min; Detection predominant wavelength is 340nm, and commplementary wave length is 405nm.
embodiment 3
Mensuration kit is composed as follows:
1. reagent R1 is:
2. reagent R2 is:
Various compositions can add under room temperature successively, or add simultaneously, or packaging preparation immediately again before detecting separately respectively.
3. reagent detection ratio is 4: 1
The CRE detection kit that the present embodiment is described, is applicable to various types of full automatic biochemical apparatus, and taking Hitachi's 7170 full automatic biochemical apparatus as example, it operates as table 1.Analytical approach: Two point end assay, the consumption of reagent R1, R2 is respectively 240ul and 60ul, sample size 3.5ul; 240ul reagent R1 adds 3.5ul sample to hatch 5min in 37 DEG C, adds 60ul R2, hatches 1.5min for 37 DEG C, monitors continuously the variation of 180s absorbance, calculates △ A/min; Detection predominant wavelength is 340nm, and commplementary wave length is 405nm.
embodiment 4
Mensuration kit is composed as follows:
1. reagent R1 is:
2. reagent R2 is:
Various compositions can add under room temperature successively, or add simultaneously, or packaging preparation immediately again before detecting separately respectively.
3. reagent detection ratio is 4: 1
The CRE detection kit that the present embodiment is described, is applicable to various types of full automatic biochemical apparatus, and taking Hitachi's 7170 full automatic biochemical apparatus as example, it operates as table 1.Analytical approach: Two point end assay, the consumption of reagent R1, R2 is respectively 240ul and 60ul, sample size 3.5ul; 240ul reagent R1 adds 3.5ul sample to hatch 5min in 37 DEG C, adds 60ul R2, hatches 1.5min for 37 DEG C, monitors continuously the variation of 180s absorbance, calculates △ A/min; Detection predominant wavelength is 340nm, and commplementary wave length is 405nm.
embodiment 5
Mensuration kit is composed as follows:
1. reagent R1 is:
2. reagent R2 is:
Various compositions can add under room temperature successively, or add simultaneously, or packaging preparation immediately again before detecting separately respectively.
3. reagent detection ratio is 4: 1
The CRE detection kit that the present embodiment is described, is applicable to various types of full automatic biochemical apparatus, and taking Hitachi's 7170 full automatic biochemical apparatus as example, it operates as table 1.Analytical approach: Two point end assay, the consumption of reagent R1, R2 is respectively 240ul and 60ul, sample size 3.5ul; 240ul reagent R1 adds 3.5ul sample to hatch 5min in 37 DEG C, adds 60ul R2, hatches 1.5min for 37 DEG C, monitors continuously the variation of 180s absorbance, calculates △ A/min; Detection predominant wavelength is 340nm, and commplementary wave length is 405nm.
the correlation test of embodiment 6, detection reagent
Use the Urea reagent of this law invention reagent (specifically filling a prescription with embodiment 1) and contrast agents Beckman company, adopt automatic 7170 automatic clinical chemistry analyzers to measure by each autoregressive parameter 50 parts of human serums (comprising normal and monstrosity) simultaneously, measured value is carried out to correlation analysis.According to above-mentioned " table 1 " in parameter measure measurement result and see Fig. 2, X, Y-axis is measured value (the content mmol/L of Urea), is found out by the result of Fig. 2, the relevant of two kinds of reagent is R
2=0.9977, regression equation is y=1.1598x-0.7773.It is good that result shows that this reagent and import reagent are measured patients serum's correlativity, has good specificity and accuracy.
In addition, above experiment is that 7170 full automatic biochemical apparatus that adopt Hitachi, Ltd to manufacture carry out, but reagent of the present invention is not limited to above-mentioned instrument, is also applicable to other full-automatic or semi-automatic biochemical analyzers.
embodiment 7, replica test
This experiment purpose is to detect reagent repeatability.Adopt embodiment 1 reagent, contrast agents (Nanjing Australia woods biology), standard items, blank solution (being generally normal saline solution and Purified Water), normal human serum sample.
Machine: Hitachi's 7170 automatic biochemistry analyzers.
Operation steps: test 10 times serum sample, calculate average and SD numerical value, try to achieve CV numerical value.
Result is resolved: according to detecting data, calculate CV numerical value, CV numerical value more approaches 0, represents that repeatability is better.
Table 2 result shows, the coefficient of variation CV=0.70% of reagent detection Urea of the present invention, and Nanjing Australia woods reagent detects the coefficient of variation CV=3.37% of Urea, and the CV value that reagent of the present invention detects reagent is less than Nanjing Australia woods reagent, shows that reagent of the present invention is reproducible.
The replica test of embodiment 2～5 reagent is the same, and testing result, substantially with embodiment 1, shows that reagent of the present invention has good repeatability.
Table 2
embodiment 8, accuracy experiment
This experiment purpose is to detect reagent accuracy.
Operation steps: test respectively the high and low value quality controlled serum of Roche 3 times, calculate average and the relative deviation with quality-control product target value.
Result is resolved: according to detecting data, calculate relative deviation, the absolute value of deviation is less, represents that accuracy is higher.The demonstration of table 3 result, reagent of the present invention (embodiment 1 reagent) is measured the high and low value determination of serum of the multinomial biochemistry quality control of Roche mean value and is respectively 6.83mmol/L, 20.58mmol/L, is respectively 0.24% and 2.92% with the relative deviation of target value; Nanjing Australia woods reagent is measured mean value and is respectively 6.72mmol/L, 21.51mmol/L, is respectively-1.37% and 7.55% with the relative deviation of target value.The accuracy that shows reagent of the present invention is high.
The accuracy test of embodiment 2～5 reagent is the same, and testing result, substantially with embodiment 1, shows that reagent of the present invention has good accuracy.
Table 3
in embodiment 9, invention reagent and R1, do not add the survey Serum experiments of glutamte dehydrogenase and NADH reagent
This experiment purpose is to detect reagent to eliminate serum endogenous ammonia interference effect.
Operation steps: use and do not add the reagent of glutamte dehydrogenase and NADH and the Urea reagent of contrast agents Beckman company in reagent of the present invention (embodiment 1 reagent), R1, adopt automatic 7170 automatic clinical chemistry analyzers to measure by each autoregressive parameter 20 parts of human serums (comprising normal and monstrosity) simultaneously, measured value is carried out to correlation analysis.
Fig. 3 is reagent of the present invention and Beckman company reagent serum correlativity schematic diagram, and what wherein X-axis represented is patients serum's result that reagent of the present invention is measured, and what Y-axis represented is patients serum's result that Beckman company reagent is measured, coefficient R
2=0.9959, regression equation is y=1.0394x-0.1043, and both correlativitys are good as shown in Figure 3.
Fig. 4 is that in kit of the present invention, R1 does not add glutamte dehydrogenase and NADH reagent and Beckman company reagent serum correlativity schematic diagram, what wherein X-axis represented is patients serum's result that R1 does not add glutamte dehydrogenase and NADH reagent mensuration, what Y-axis represented is patients serum's result that Beckman company reagent is measured, coefficient R
2=0.7881, regression equation is y=0.9147x+0.2858, and both correlativitys are not good as shown in Figure 4.
Table 4 result shows, reagent of the present invention and the reagent serum correlativity R of Beckman company
2in=0.9959, R1, do not add reagent and the reagent serum correlativity R of Beckman company of glutamte dehydrogenase and two kinds of compositions of NADH
2=0.7881, show to add in R1 glutamte dehydrogenase and NADH can effectively eliminate endogenous ammonia and disturb, good with Beckman company reagent serum correlativity.
Table 4
embodiment 10, enzymatic protective reagent detect the impact of reagent on urea
Detect reagent of the present invention, add conventional enzymatic protective reagent BSA reagent and do not add the impact of enzymatic protective reagent reagent on urease.
Table 5 represents reagent of the present invention and does not add the variation of urease activity in the reagent 1 year of enzymatic protective reagent dimethyl sulfoxide (DMSO); Table 5 result shows; the activity that does not add protectant reagent urease within a year drops to 22% from 100%; add the enzymatic activity in the reagent 1 year of protective agent BSA to decline 40%, and add the enzymatic activity in the reagent 1 year of protective agent dimethyl sulfoxide (DMSO) only to decline 22%.After 1 year with the reagent of protective agent dimethyl sulfoxide (DMSO) compared with not adding protectant reagent, the activity of urease exceeds 56%, adds the reagent of dimethyl sulfoxide (DMSO) to compare with the reagent that adds BSA, enzymatic activity promote 28%.Result shows that dimethyl sulfoxide (DMSO) is very strong to the protective effect of enzyme, and more effective than BSA, As time goes on, more obvious to the protective effect meeting of enzyme, thereby improves the stability of reagent of the present invention.
Table 5
Above specific embodiments of the invention are described.It will be appreciated that, the present invention is not limited to above-mentioned specific implementations, and those skilled in the art can make various distortion or amendment within the scope of the claims, and this does not affect flesh and blood of the present invention.
Claims (10)
1. a detection kit that is not subject to the mensuration urea content of serum endogenous ammonia interference, is characterized in that, comprises reagent R1 and reagent R2; Damping fluid, 1～80KU/L glutamte dehydrogenase, 0.5～2KU/L NADH, 1～100mmol/L α-ketoglutaric acid, 1 ‰～5 ‰ surfactant and 0.1～1g/L antiseptic that described reagent R1 contains 1～500mmol/L pH=3.0～8.0; Damping fluid, 1～60KU/L urease, 0.5～2KU/L NADH, 1～100mmol/L α-ketoglutaric acid, 10～500mmol/L sodium chloride, 1～50mmol/L enzymatic protective reagent and 0.1～1g/L antiseptic that described reagent R2 contains 1～500mmol/L pH=3.0～8.0.
2. detection kit according to claim 1, it is characterized in that damping fluid, 20～60KU/L glutamte dehydrogenase, 1～1.5KU/L NADH, 50～100mmol/L α-ketoglutaric acid, 1 ‰～5 ‰ surfactant and 0.2～0.8g/L antiseptic that described reagent R1 contains 100～300mmol/L pH=5.0～8.0; Damping fluid, 10～30KU/L urease, 1～1.5KU/L NADH, 50～100mmol/L α-ketoglutaric acid, 100～300mmol/L sodium chloride, 10～40mmol/L enzymatic protective reagent and 0.2～0.8g/L antiseptic that described reagent R2 contains 100～300mmol/L pH=5.0～8.0.
3. detection kit according to claim 1, is characterized in that, the damping fluid comprising in described reagent R1, R2 is selected from respectively TRIS, MOPS, HEPES or BICN.
4. detection kit according to claim 3, is characterized in that, the damping fluid comprising in described reagent R1, R2 is TRIS.
5. detection kit according to claim 1, is characterized in that, described enzymatic protective reagent is dimethyl sulfoxide (DMSO).
6. detection kit according to claim 1, is characterized in that, described surfactant is polysorbas20, Tween 80 or triton x-100.
7. detection kit according to claim 1, is characterized in that, the antiseptic comprising in described reagent R1, R2 is selected from respectively sodium azide, proclin300, polylysine or sodium acetate.
8. one kind according to the using method of the detection kit described in any one in claim 1～7, it is characterized in that, reagent R1 adds after sample 37 DEG C to hatch 5 minutes, add again reagent R2, hatch 1.5 minutes for 37 DEG C, the monitoring absorbance rate of change of 3 minutes, compares with urea content and the absorbance relation curve of Urea standard items continuously, obtains the urea content of sample.
9. the using method of detection kit according to claim 8, is characterized in that, the predominant wavelength that described monitoring adopts is 340nm, and commplementary wave length is 405nm.
10. the using method of detection kit according to claim 8, is characterized in that, the volume ratio of described reagent R1 and reagent R2 is 4: 1.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107014969A (en) * | 2016-01-27 | 2017-08-04 | 北京凯茂正奇科技有限公司 | Detect the agent prescription of urea and ammonia content |
| CN107064483A (en) * | 2017-04-26 | 2017-08-18 | 上海健康医学院 | A kind of double item rapid assay methods of serum urea nitrogen creatinine |
| CN107121325A (en) * | 2017-05-31 | 2017-09-01 | 江西乐成生物医疗有限公司 | A kind of detection method that diagnosis is damaged for early hepatocyte |
| CN109374884A (en) * | 2018-12-24 | 2019-02-22 | 四川沃文特生物技术有限公司 | A kind of PCT concentration detection kit and preparation method thereof |
| CN109387645A (en) * | 2018-10-19 | 2019-02-26 | 蓝怡科技集团股份有限公司 | A kind of serum urea measurement reagent and its application |
| CN112126674A (en) * | 2020-08-18 | 2020-12-25 | 兰州百源基因技术有限公司 | A kind of urea detection kit, its preparation method and use method |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4189536A (en) * | 1977-11-09 | 1980-02-19 | Hycel, Inc. | Reagent systems, enzymatic assays and compositions therefor |
| EP0293732A2 (en) * | 1987-06-03 | 1988-12-07 | Abbott Laboratories | Simultaneous assay for glucose and urea |
| CN101514357A (en) * | 2009-04-01 | 2009-08-26 | 北京瑞正善达生物工程技术有限公司 | Clinic in vitro potassium determination reagent by enzyme method |
| CN101571488A (en) * | 2008-04-28 | 2009-11-04 | 北京华大吉比爱生物技术有限公司 | Method and kit for measuring urea nitrogen |
| CN102928468A (en) * | 2012-09-14 | 2013-02-13 | 北京体育大学 | Whole blood urea biosensing test paper |
| CN103276052A (en) * | 2013-05-24 | 2013-09-04 | 宁波美康生物科技股份有限公司 | Urea nitrogen detection reagent |
-
2014
- 2014-08-14 CN CN201410401509.7A patent/CN104198421A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4189536A (en) * | 1977-11-09 | 1980-02-19 | Hycel, Inc. | Reagent systems, enzymatic assays and compositions therefor |
| EP0293732A2 (en) * | 1987-06-03 | 1988-12-07 | Abbott Laboratories | Simultaneous assay for glucose and urea |
| EP0293732A3 (en) * | 1987-06-03 | 1991-03-06 | Abbott Laboratories | Simultaneous assay for glucose and urea |
| CN101571488A (en) * | 2008-04-28 | 2009-11-04 | 北京华大吉比爱生物技术有限公司 | Method and kit for measuring urea nitrogen |
| CN101514357A (en) * | 2009-04-01 | 2009-08-26 | 北京瑞正善达生物工程技术有限公司 | Clinic in vitro potassium determination reagent by enzyme method |
| CN102928468A (en) * | 2012-09-14 | 2013-02-13 | 北京体育大学 | Whole blood urea biosensing test paper |
| CN103276052A (en) * | 2013-05-24 | 2013-09-04 | 宁波美康生物科技股份有限公司 | Urea nitrogen detection reagent |
Non-Patent Citations (2)
| Title |
|---|
| CRTER_NRR: "冻存保护剂", 《百度百科》 * |
| 蒋兴亮 等: "脲酶和亮氨酸脱氢酶偶联法动力学测定血清或尿液中尿素", 《临床检验杂志》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107014969A (en) * | 2016-01-27 | 2017-08-04 | 北京凯茂正奇科技有限公司 | Detect the agent prescription of urea and ammonia content |
| CN107064483A (en) * | 2017-04-26 | 2017-08-18 | 上海健康医学院 | A kind of double item rapid assay methods of serum urea nitrogen creatinine |
| CN107121325A (en) * | 2017-05-31 | 2017-09-01 | 江西乐成生物医疗有限公司 | A kind of detection method that diagnosis is damaged for early hepatocyte |
| CN109387645A (en) * | 2018-10-19 | 2019-02-26 | 蓝怡科技集团股份有限公司 | A kind of serum urea measurement reagent and its application |
| CN109374884A (en) * | 2018-12-24 | 2019-02-22 | 四川沃文特生物技术有限公司 | A kind of PCT concentration detection kit and preparation method thereof |
| CN112126674A (en) * | 2020-08-18 | 2020-12-25 | 兰州百源基因技术有限公司 | A kind of urea detection kit, its preparation method and use method |
| CN112126674B (en) * | 2020-08-18 | 2024-02-06 | 兰州百源基因技术有限公司 | A urea detection kit, its preparation method and use method |
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