CN113655006B - Urinary system knot Dan Chengdan risk factor detection and test system - Google Patents
Urinary system knot Dan Chengdan risk factor detection and test system Download PDFInfo
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- 238000001514 detection method Methods 0.000 title claims abstract description 122
- 230000002485 urinary effect Effects 0.000 title claims abstract description 69
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 216
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims abstract description 132
- 210000002700 urine Anatomy 0.000 claims abstract description 71
- 239000011777 magnesium Substances 0.000 claims abstract description 32
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 claims abstract description 31
- 229910052749 magnesium Inorganic materials 0.000 claims abstract description 31
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims abstract description 30
- 239000011575 calcium Substances 0.000 claims abstract description 30
- 229910052791 calcium Inorganic materials 0.000 claims abstract description 30
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims abstract description 29
- 239000011574 phosphorus Substances 0.000 claims abstract description 29
- 229910052698 phosphorus Inorganic materials 0.000 claims abstract description 29
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 claims abstract description 26
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 claims abstract description 26
- 229940116269 uric acid Drugs 0.000 claims abstract description 26
- 239000002253 acid Substances 0.000 claims abstract description 11
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 claims description 101
- 235000006408 oxalic acid Nutrition 0.000 claims description 38
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 claims description 28
- 239000000243 solution Substances 0.000 claims description 25
- 102000003992 Peroxidases Human genes 0.000 claims description 21
- 108040007629 peroxidase activity proteins Proteins 0.000 claims description 21
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 18
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 claims description 18
- RLFWWDJHLFCNIJ-UHFFFAOYSA-N 4-aminoantipyrine Chemical compound CN1C(C)=C(N)C(=O)N1C1=CC=CC=C1 RLFWWDJHLFCNIJ-UHFFFAOYSA-N 0.000 claims description 14
- 102000004316 Oxidoreductases Human genes 0.000 claims description 14
- 108090000854 Oxidoreductases Proteins 0.000 claims description 14
- PHOLIFLKGONSGY-CSKARUKUSA-N (e)-(3-methyl-1,3-benzothiazol-2-ylidene)hydrazine Chemical compound C1=CC=C2S\C(=N\N)N(C)C2=C1 PHOLIFLKGONSGY-CSKARUKUSA-N 0.000 claims description 12
- 239000007853 buffer solution Substances 0.000 claims description 12
- -1 azo arsenic III Chemical compound 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 108010036824 Citrate (pro-3S)-lyase Proteins 0.000 claims description 9
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 9
- 102000013460 Malate Dehydrogenase Human genes 0.000 claims description 9
- 108010026217 Malate Dehydrogenase Proteins 0.000 claims description 9
- JLTDJTHDQAWBAV-UHFFFAOYSA-N N,N-dimethylaniline Chemical compound CN(C)C1=CC=CC=C1 JLTDJTHDQAWBAV-UHFFFAOYSA-N 0.000 claims description 9
- 239000011780 sodium chloride Substances 0.000 claims description 9
- 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 8
- 239000007990 PIPES buffer Substances 0.000 claims description 8
- 239000012452 mother liquor Substances 0.000 claims description 8
- 239000000843 powder Substances 0.000 claims description 8
- 108010092464 Urate Oxidase Proteins 0.000 claims description 7
- 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 description 7
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 claims description 7
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 7
- 239000010452 phosphate Substances 0.000 claims description 7
- 108010024957 Ascorbate Oxidase Proteins 0.000 claims description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 6
- 229910019142 PO4 Inorganic materials 0.000 claims description 6
- 239000007983 Tris buffer Substances 0.000 claims description 6
- APUPEJJSWDHEBO-UHFFFAOYSA-P ammonium molybdate Chemical compound [NH4+].[NH4+].[O-][Mo]([O-])(=O)=O APUPEJJSWDHEBO-UHFFFAOYSA-P 0.000 claims description 6
- 239000011609 ammonium molybdate Substances 0.000 claims description 6
- 235000018660 ammonium molybdate Nutrition 0.000 claims description 6
- 229940010552 ammonium molybdate Drugs 0.000 claims description 6
- DEFVIWRASFVYLL-UHFFFAOYSA-N ethylene glycol bis(2-aminoethyl)tetraacetic acid Chemical compound OC(=O)CN(CC(O)=O)CCOCCOCCN(CC(O)=O)CC(O)=O DEFVIWRASFVYLL-UHFFFAOYSA-N 0.000 claims description 6
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims description 6
- 210000001635 urinary tract Anatomy 0.000 claims description 5
- 239000007864 aqueous solution Substances 0.000 claims description 4
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 claims description 3
- 102000004190 Enzymes Human genes 0.000 claims description 3
- 108090000790 Enzymes Proteins 0.000 claims description 3
- 239000004471 Glycine Substances 0.000 claims description 3
- 239000000872 buffer Substances 0.000 claims description 3
- BVKZGUZCCUSVTD-UHFFFAOYSA-N carbonic acid Chemical compound OC(O)=O BVKZGUZCCUSVTD-UHFFFAOYSA-N 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 2
- 238000000034 method Methods 0.000 abstract description 8
- 238000004364 calculation method Methods 0.000 abstract description 2
- 239000000306 component Substances 0.000 description 12
- 208000009911 Urinary Calculi Diseases 0.000 description 9
- 238000011088 calibration curve Methods 0.000 description 8
- 238000002835 absorbance Methods 0.000 description 6
- 238000005259 measurement Methods 0.000 description 6
- 239000007850 fluorescent dye Substances 0.000 description 5
- 230000008901 benefit Effects 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 230000000927 lithogenic effect Effects 0.000 description 4
- 230000035945 sensitivity Effects 0.000 description 4
- 206010007027 Calculus urinary Diseases 0.000 description 3
- 208000015924 Lithiasis Diseases 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 238000011282 treatment Methods 0.000 description 3
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- SYXIQGAAYVTLEO-UHFFFAOYSA-N [P].N1C(=O)NC(=O)C2=C1NC(=O)N2 Chemical compound [P].N1C(=O)NC(=O)C2=C1NC(=O)N2 SYXIQGAAYVTLEO-UHFFFAOYSA-N 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- POJWUDADGALRAB-UHFFFAOYSA-N allantoin Chemical compound NC(=O)NC1NC(=O)NC1=O POJWUDADGALRAB-UHFFFAOYSA-N 0.000 description 2
- 208000035269 cancer or benign tumor Diseases 0.000 description 2
- 238000006555 catalytic reaction Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000007979 citrate buffer Substances 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 238000000556 factor analysis Methods 0.000 description 2
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- 238000011534 incubation Methods 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 230000006371 metabolic abnormality Effects 0.000 description 2
- YBSLUBPQYDULIZ-UHFFFAOYSA-N oxalic acid;urea Chemical compound NC(N)=O.OC(=O)C(O)=O YBSLUBPQYDULIZ-UHFFFAOYSA-N 0.000 description 2
- 230000003252 repetitive effect Effects 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000004575 stone Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- YQOFSYQCZHXAOZ-UHFFFAOYSA-N 2-hydroxypropane-1,2,3-tricarboxylic acid;2-oxobutanedioic acid Chemical compound OC(=O)CC(=O)C(O)=O.OC(=O)CC(O)(C(O)=O)CC(O)=O YQOFSYQCZHXAOZ-UHFFFAOYSA-N 0.000 description 1
- POJWUDADGALRAB-PVQJCKRUSA-N Allantoin Natural products NC(=O)N[C@@H]1NC(=O)NC1=O POJWUDADGALRAB-PVQJCKRUSA-N 0.000 description 1
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- 208000000913 Kidney Calculi Diseases 0.000 description 1
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 description 1
- 206010029148 Nephrolithiasis Diseases 0.000 description 1
- 159000000021 acetate salts Chemical class 0.000 description 1
- 229960000458 allantoin Drugs 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 239000008358 core component Substances 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 235000019007 dietary guidelines Nutrition 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 150000002466 imines Chemical class 0.000 description 1
- 239000002932 luster Substances 0.000 description 1
- 229910001425 magnesium ion Inorganic materials 0.000 description 1
- 229910000157 magnesium phosphate Inorganic materials 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 238000012502 risk assessment Methods 0.000 description 1
- 238000005096 rolling process Methods 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 208000008281 urolithiasis Diseases 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/27—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using photo-electric detection ; circuits for computing concentration
- G01N21/274—Calibration, base line adjustment, drift correction
- G01N21/278—Constitution of standards
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
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- Spectroscopy & Molecular Physics (AREA)
- Engineering & Computer Science (AREA)
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- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a urinary system Dan Chengdan risk factor detection and test system, which comprises a matched instrument and a kit, wherein a plurality of reagents are arranged in the kit, and comprise a urooxalic acid detection reagent, a calcium urine detection reagent, a citric acid urine detection reagent, a magnesium urine detection reagent, a phosphorus urine detection reagent and a uric acid urine detection reagent. The invention integrates a series of reagents (the urooxalic acid detection reagent, the calcium urine detection reagent, the citric acid urine detection reagent, the magnesium urine detection reagent, the phosphorus urine detection reagent and the uric acid detection reagent) on one instrument for detection, and is more convenient. The method is suitable for high-flux detection, is simple to operate, only needs to put the series of reagents into a reagent bin, then puts a urine sample into a sample bin, automatically detects the urine sample by an instrument, automatically outputs a result, and does not need manual calculation.
Description
Technical Field
The invention relates to the technical field of detection kits, in particular to a urinary system Dan Chengdan risk factor detection and test system.
Background
Urinary calculi are a common disease in China, and generally show that the incidence rate is higher in the southern area than in the northern area. One study taught in Zeng Guohua of our country in 2015 shows that the prevalence of kidney stones in adult China is 5.8%, with a prevalence of 6.5% for men and 5.1% for women.
Although with the improvement of the technological level, the treatment mode and effect of the urinary calculi are obviously improved. However, the high recurrence rate of urinary calculi, the residual rate of calculi and the regeneration rate of residual calculi are also high. Metabolic abnormalities are closely related to the formation of urinary calculi. By assessing the metabolism of urinary tract lithiasis patients, it has become an important method for revealing and diagnosing the cause of lithiasis.
The most interesting and effective metabolic abnormality examination items of urinary system lithology patients at present abroad comprise 24-hour urinary lithogenesis risk factor analysis and the like. The known urinary lithogenic risk factors are mainly high oxalic acid urine, high calcium urine, low citric acid urine, low magnesium urine, high phosphorus urine, high uric acid urine and the like. The combined detection of these lithogenic risk factors helps to aid in the diagnosis of the composition of urinary stones, thereby guiding clinical treatment, and to administer dietary guidelines or corresponding medication based on the respective stone type, preventing and reducing recurrence of stones. Thus, urine lithogenic risk factor analysis is important for a lithiasis patient to assess lithogenic risk factors/metabolic factors.
Currently, the detection of urinary oxalic acid, urinary calcium, urinary citric acid, urinary magnesium, urinary phosphorus, urinary uric acid, and the like has been widely performed in many developed and developing countries. But is not developed on a large scale in our country, which is related to the inability of most hospitals to directly detect these Dan Weixian factors. The main difficulty is that the projects need to be carried out on different instruments and equipment, and although calcium, magnesium, phosphorus and uric acid can be carried out on a conventional biochemical analyzer, the detection of oxalic acid and citric acid is still carried out on an ion chromatograph at present, and the ion chromatograph equipment is expensive, time-consuming to operate and cannot be used for conventional detection. Detection on different instruments and equipment is complex in operation, time-consuming, difficult to popularize and incapable of realizing high-flux detection. Therefore, most hospitals have not developed.
Disclosure of Invention
The invention aims to provide a urinary calculus forming risk factor detection and test system, which can realize detection of urinary oxalic acid, urinary calcium, urinary citric acid, urinary magnesium, urinary phosphorus and urinary uric acid through one kit and has the advantage of convenient operation.
The invention is realized by the following technical scheme:
the urinary system Dan Chengdan risk factor detection test system comprises a matched instrument and a kit, wherein a plurality of reagents are arranged in the kit, and comprise a urinary oxalic acid detection reagent, a urinary calcium detection reagent, a urinary citric acid detection reagent, a urinary magnesium detection reagent, a urinary phosphorus detection reagent and a urinary uric acid detection reagent;
the uric acid detection reagent comprises a first R1 reagent and a first R2 reagent;
the first R1 reagent comprises the following components:
citric acid, DMAB and MBTH, wherein the concentration of the citric acid in the first R1 reagent is 0.01-1M, and the concentration of the DMAB in the first R1 reagent is 0.1-0.5mM; the concentration of MBTH in the first R1 reagent is 0.01-0.5mM;
the first R2 reagent comprises the following components:
buffer solution with pH value of 3.6-6, oxalic acid oxidase and peroxidase; the concentration of the buffer solution in the first R2 reagent is 0.01-0.5M, the concentration of the oxalic acid oxidase in the first R2 reagent is more than or equal to 50U/L, and the concentration of the peroxidase in the first R2 reagent is more than or equal to 5000U/L;
the first R2 reagent is in a dry powder state, and is re-dissolved by water when in use;
the urine citric acid detection reagent comprises a second R1 reagent and a second R2 reagent;
the second R1 reagent comprises the following components: tris buffer solution, NADH and sodium azide, wherein the concentration of the Tris buffer solution is 40mmol/L, the concentration of NADH is 0.4mmol/L, and the concentration of sodium azide is 0.95g/L;
the second R2 reagent includes an R2a reagent and an R2b reagent:
the R2a reagent comprises PIPES and sodium azide, the PIPES concentration is 600mmol/L, and the sodium azide concentration is 0.95g/L;
the R2b reagent comprises malate dehydrogenase and citrate lyase, and the concentration of the malate dehydrogenase is more than 40KU/L; the concentration of the citrate lyase is more than 1KU/L;
the R2b reagent is in a dry powder state, and R2a and R2b are mixed to obtain a reagent second R2 reagent when in use;
the urine calcium detection reagent comprises an R reagent, wherein the R reagent comprises the following components;
azo arsenic III and imidazole; the concentration of azo arsenic III is 0.2mmol/L, and the concentration of imidazole is 75mmol/L.
The magnesium urine detection reagent comprises a third R1 reagent, a third R2 reagent and a third R2 reagent:
the third R1 reagent comprises the following components:
carbonate buffer and glycol ether diamine tetraacetic acid; the concentration of the carbonic acid buffer solution is 0.1mol/L, and the concentration of the glycol ether diamine tetraacetic acid is 0.1mmol/L;
the third R2 reagent comprises dimethylaniline blue and glycine; the concentration of the dimethylaniline blue is 0.5mmol/L; glycine concentration was 25mmol/L;
the urine phosphorus detection reagent comprises a fourth R1 reagent and a fourth R2 reagent;
the fourth R1 reagent comprises sodium chloride with the concentration of 154mmol/L;
the fourth R2 reagent comprises sodium chloride and ammonium molybdate; the concentration of sodium chloride is 154mmol/L; the concentration of ammonium molybdate is 3.5mmol/L;
the urine acid detection reagent comprises the following components:
phosphate, uricase, peroxidase, 4-aminoantipyrine and ascorbate oxidase, wherein the phosphate concentration is 100mmol/L; uricase concentration is greater than 0.12U/ml; the concentration of peroxidase is more than or equal to 1U/mL;4 aminoantipyrine concentration was 0.5mmol/L; the concentration of ascorbate oxidase is greater than 5U/mL.
The invention has the conception that:
a series of reagents (an urooxalic acid detection reagent, a calcium urine detection reagent, a citric acid urine detection reagent, a magnesium urine detection reagent, a phosphorus urine detection reagent and an uric acid detection reagent) are integrated on one instrument for detection, so that the detection is more convenient. The method is suitable for high-flux detection, is simple to operate, only needs to put the series of reagents into a reagent bin, then puts a urine sample into a sample bin, automatically detects the urine sample by an instrument, automatically outputs a result, and does not need manual calculation.
Wherein, the measuring principle of the urooxalic acid is as follows: the oxalic acid is subjected to coupling reaction under the action of oxalic acid oxidase and peroxidase to generate a colored compound, and the concentration of oxalic acid can be calculated according to the absorbance of the compound.
The oxalic acid detection reagent has the core components of oxalic acid oxidase and peroxidase, but the optimal pH values of the oxalic acid oxidase and the peroxidase are different, the invention ensures higher activity of the two enzymes by reasonably designing the formula of the R2 reagent, thereby achieving the characteristic of high sensitivity, and simultaneously, the whole reagent kit has high sensitivity and high detection limit by reasonably designing the formula of the R1 reagent, so that the reagent kit can be suitable for detecting oxalic acid in urine.
The method has higher detection limit of 1.8-180mg/L, and can directly measure the oxalic acid content in urine without diluting the sample.
In the existing oxalic acid measurement, most of detection kits have low detection limit, basically 9-45mg/L, and are only suitable for detecting the oxalic acid content in blood, and when the kit is used for detecting the oxalic acid content in urine, a sample needs to be diluted, so that the accuracy of a detection result is reduced.
In addition, the reagent does not contain reagents such as fluorescent dye, the fluorescent dye is not stable in the air, the fluorescent dye is used as soon as possible after being started, the fluorescent dye is unstable in the presence of a reducing agent, the anti-interference capability of the reagent is weak, and the reagent avoids pollution of the fluorescent dye to the environment. The reagent variety is simple, and only two reagents R1 and R2 are adopted. And the incubation is not needed, the time waste caused by incubation is reduced, the detection can be directly carried out on the machine, the calibration mode is simple, and the calibration is not needed for each detection. The operation is simple and quick, the result can be obtained after 10 minutes after the machine is started, the time is quick, the method is more suitable for high-flux detection and instant detection, and the method is more suitable for mass operation of medical laboratories and clinical laboratory in hospitals.
Wherein, the measurement principle of the urine citric acid is as follows: the concentration of citric acid can be calculated by inversely proportional to the change value of absorbance of the enzymatic reaction substrate NADH under the action of the citrate lyase and the malate dehydrogenase.
Citric acid-oxaloacetic acid + acetate salt
Wherein, the measurement principle of the urine acid is as follows: uric Acid (UA) is oxidized to generate allantoin and H2O2 under the catalysis of uricase, and the H2O2, 4-aminoantipyrine and 3, 5-dioxo-2-hydroxybenzenesulfonic acid generate colored substances (rolling imine compounds) under the catalysis of peroxidase, wherein the color and luster of the colored substances are in direct proportion to the concentration of UA in a sample.
Wherein, the measurement principle of urine calcium is as follows: the calcium can react with azo arsenic III (ArsenazoIII) in the reagent, and the generated colored complex can be measured by a colorimetry method.
Wherein, the measurement principle of the urine magnesium is as follows: magnesium (Mg) and the dimethylaniline blue are combined to generate a purple water-soluble neoplasm compound, the purple water-soluble neoplasm compound has maximum absorbance at 520nm, simultaneously, the dimethylaniline blue is used as a substrate to be continuously reduced, the absorbance of the dimethylaniline blue is also continuously reduced, and the concentration of magnesium in a sample can be obtained by measuring the absorbance reduction of the substrate.
Wherein, the measurement principle of urine phosphorus is: by adopting a phosphomolybdate complex direct analysis method, unreduced phosphomolybdate complex generated by the reaction is in direct proportion to the content of phosphorus in a sample. The concentration of inorganic phosphorus in the sample can be calculated by measuring the change of absorbance value at 340 nm.
The kit also comprises a standard urooxalic acid product, a standard calcium uronate product, a standard citric acid uronate product, a standard magnesium uronate product, a standard phosphorus uronate product and a standard uric acid.
Further, the preparation process of the first R1 reagent is as follows:
dissolving DMAB in methanol to prepare DMAB mother liquor, dissolving MBTH in water to prepare MBTH mother liquor, dissolving citric acid in water to prepare citric acid solution, and mixing the DMAB mother liquor, the MBTH mother liquor and the citric acid solution in proportion to obtain the first R1 reagent.
Further, the preparation process of the first R2 reagent is as follows:
the oxalic acid oxidase is redissolved with water to form a solution and stored at 2-8 ℃, the peroxidase is redissolved with water to form a solution and stored at 2-8 ℃, and the buffer solution, the oxalic acid oxidase aqueous solution and the peroxidase aqueous solution are mixed according to a proportion to obtain a first R2 reagent.
Further, the preparation process of the second R2 reagent is as follows:
PIPES and sodium azide are mixed according to a proportion to form R2a, malate dehydrogenase is redissolved to form a solution, citrate lyase is redissolved to form a solution, and the two enzymes are mixed according to a proportion to form R2b and are freeze-dried to form dry powder.
Further, oxalic acid detection reagent, calcium urinate detection reagent, citric acid urinate detection reagent, magnesium urinate detection reagent, phosphorus urinate detection reagent and uric acid detection reagent are arranged in a partitioned manner.
Further, the instrument includes BST300 and BA200.
The urinary calculus risk factor detection and test system is placed in a reagent disk corresponding to an instrument, a collected specimen is placed in a corresponding specimen position, the instrument automatically detects the specimen, and the result is automatically calculated.
The BST300 full-automatic specific protein analyzer comprises an instrument external structure, a sample adding system, a cleaning system, a stirring system, a reaction system, an optical system and the like; the BA200 full-automatic specific protein analyzer comprises an instrument external structure, a sample adding system, a cleaning system, a stirring system, a reaction system, an optical system and the like.
The specific detection steps are as follows:
1) Calibration procedure: and (3) placing the urooxalic acid standard, the calcium urine standard, the citric acid urine standard, the magnesium urine standard, the phosphorus urine standard and the uric acid urine standard at a designated position of an instrument (BioSystems A400/BA200 full-automatic specific protein analyzer) for calibration, so as to obtain a calibration curve.
2) Sample detection flow:
and (3) placing the collected clinical urine sample, the urinary oxalic acid detection reagent, the urinary calcium detection reagent, the urinary citric acid detection reagent, the urinary magnesium detection reagent, the urinary phosphorus detection reagent and the urinary uric acid detection reagent at corresponding positions of an instrument, starting the instrument to directly measure the contents of urinary oxalic acid, urinary calcium, urinary citric acid, urinary magnesium, urinary phosphorus and urinary uric acid, and obtaining a first result about 8 minutes.
Compared with the prior art, the invention has the following advantages and beneficial effects:
1. the invention can realize the detection of the urinary oxalic acid, the urinary calcium, the urinary citric acid, the urinary magnesium, the urinary phosphorus and the urinary uric acid by one kit, has the advantage of convenient operation, and provides basis for risk assessment, diagnosis, treatment and relapse prevention of urinary calculi.
2. The oxalic acid detection reagent of the invention enables oxalic acid oxidase and peroxidase to reach higher activity under proper PH condition, and improves detection sensitivity, and the linear range of the oxalic acid detection kit measured by the invention is 1.8-180mmol/L.
3. The kit can be used on a matched instrument, achieves the aim of directly connecting with a machine, and is more convenient to use.
4. The matched instrument comprises the BST300 and the BA200, adopts an optical system of a global patent, and has higher sensitivity and wider linear range.
5. The detection reagents in the kit are all finished products, and the client does not need to prepare related reagents.
Drawings
The accompanying drawings, which are included to provide a further understanding of embodiments of the invention and are incorporated in and constitute a part of this application, illustrate embodiments of the invention. In the drawings:
FIG. 1 is a calibration curve of urooxalic acid in example 1;
FIG. 2 is a calibration curve of UUA in example 1;
FIG. 3 is a calibration curve of urine phosphorus in example 1;
FIG. 4 is a calibration curve of calcium urine in example 1;
FIG. 5 is a calibration curve of magnesium urine in example 1;
FIG. 6 is a calibration curve of urinary citric acid in example 1.
Detailed Description
For the purpose of making apparent the objects, technical solutions and advantages of the present invention, the present invention will be further described in detail with reference to the following examples and the accompanying drawings, wherein the exemplary embodiments of the present invention and the descriptions thereof are for illustrating the present invention only and are not to be construed as limiting the present invention.
Example 1:
the urinary system Dan Chengdan risk factor detection and test system is characterized by comprising a matched instrument and a kit, wherein the kit is internally provided with a plurality of reagents, and the plurality of reagents comprise a urinary oxalic acid detection reagent, a urinary calcium detection reagent, a urinary citric acid detection reagent, a urinary magnesium detection reagent, a urinary phosphorus detection reagent and a urinary uric acid detection reagent;
the uric acid detection reagent comprises a first R1 reagent and a first R2 reagent;
the first R1 reagent comprises the following components:
citric acid, DMAB and MBTH, the concentration of citric acid in the first R1 reagent being 0.07M, the concentration of DMAB in the first R1 reagent being 0.18mM; the concentration of MBTH in R1 reagent is 0.09mM;
the first R2 reagent comprises the following components:
buffer solution with pH value of 5, oxalic acid oxidase and peroxidase; the concentration of the buffer solution in the first R2 reagent is 0.06M, the concentration of the oxalic acid oxidase in the first R2 reagent is 100U/L, and the concentration of the peroxidase in the first R2 reagent is 5000U/L; the buffer solution is a citrate buffer solution, wherein the citrate buffer solution comprises 0.06M of citric acid and 0.06M of sodium citrate according to the volume ratio of 41: 59;
the R2 reagent is in a dry powder state, and is re-dissolved in water when in use
The urine citric acid detection reagent comprises a second R1 reagent and a second R2 reagent;
the second R1 reagent comprises the following components: tris buffer solution, NADH and sodium azide, wherein the concentration of the Tris buffer solution is 40mmol/L, the concentration of NADH is 0.4mmol/L, and the concentration of sodium azide is 0.95g/L;
the second R2 reagent includes an R2a reagent and an R2b reagent:
the R2a reagent comprises PIPES and sodium azide, the PIPES concentration is 600mmol/L, and the sodium azide concentration is 0.95g/L;
the R2b reagent comprises malate dehydrogenase and citrate lyase, and the concentration of the malate dehydrogenase is more than 40KU/L; the concentration of the citrate lyase is more than 1KU/L;
the R2b reagent is in a dry powder state, and R2a and R2b are mixed to obtain a reagent second R2 reagent when in use;
the urine calcium detection reagent comprises an R reagent, wherein the R reagent comprises the following components;
azo arsenic III and imidazole; the concentration of azo arsenic III is 0.2mmol/L, and the concentration of imidazole is 75mmol/L.
The magnesium urine detection reagent comprises a third R1 reagent, a third R2 reagent and a third R2 reagent:
the third R1 reagent comprises the following components:
carbonate buffer and glycol ether diamine tetraacetic acid; the concentration of the carbonic acid buffer solution is 0.1mol/L, and the concentration of the glycol ether diamine tetraacetic acid is 0.1mmol/L;
the third R2 reagent comprises dimethylaniline blue and glycine; the concentration of the dimethylaniline blue is 0.5mmol/L; glycine concentration was 25mmol/L;
the urine phosphorus detection reagent comprises a fourth R1 reagent and a fourth R2 reagent;
the fourth R1 reagent comprises sodium chloride with the concentration of 154mmol/L;
the fourth R2 reagent comprises sodium chloride and ammonium molybdate; the concentration of sodium chloride is 154mmol/L; the concentration of ammonium molybdate is 3.5mmol/L;
the urine acid detection reagent comprises the following components:
phosphate, uricase, peroxidase, 4-aminoantipyrine and ascorbate oxidase, wherein the phosphate concentration is 100mmol/L; uricase concentration is greater than 0.12U/ml; the concentration of peroxidase is more than or equal to 1U/mL;4 aminoantipyrine concentration was 0.5mmol/L; the concentration of ascorbate oxidase is greater than 5U/mL.
In this embodiment, the oxalic acid detection reagent, the calcium urinate detection reagent, the citric acid urinate detection reagent, the magnesium urinate detection reagent, the phosphorus urinate detection reagent and the uric acid detection reagent are arranged in a partitioned manner; the instrument employs BST300.
Accuracy data of this embodiment: the oxalic acid, calcium, magnesium, phosphorus, citric acid and uric acid accuracy was measured by preparing solutions of oxalate, calcium ion, citric acid, magnesium ion, phosphate and uric acid of 90mg/L, 2.19mmol/L, 5.2mmol/L, 0.864mmol/L, 1.39mmol/L, 379 umol/L.
The test results are shown in table 1:
TABLE 1
| Accuracy of | Urea oxalic acid | Urinary calcium | Urine citric acid | Urine magnesium | Urine phosphorus | Uric acid |
| Theoretical concentration | 90 | 2.19 | 5.2 | 0.864 | 1.39 | 379 |
| 1 | 86.2 | 2.21 | 4.98 | 0.804 | 1.3 | 390 |
| 2 | 87.64 | 2.2 | 4.78 | 0.842 | 1.29 | 378 |
| 3 | 86.34 | 2.18 | 5.21 | 0.853 | 1.38 | 368 |
| 4 | 88.93 | 2.16 | 5.33 | 0.845 | 1.33 | 389 |
| 5 | 85.43 | 2.19 | 5.19 | 0.851 | 1.28 | 365 |
| 6 | 86.29 | 2.19 | 5.05 | 0.861 | 1.41 | 389 |
| 7 | 89.2 | 2.15 | 5.00 | 0.866 | 1.38 | 377 |
| 8 | 86.1 | 2.23 | 4.92 | 0.868 | 1.39 | 367 |
| 9 | 83.32 | 2.22 | 5.03 | 0.854 | 1.32 | 372 |
| 10 | 80.23 | 2.16 | 5.12 | 0.824 | 1.37 | 389 |
| Average value of | 85.97 | 2.19 | 5.06 | 0.85 | 1.35 | 378.40 |
| Relative deviation of | -4.48％ | -0.05％ | -2.67％ | -1.99％ | -3.24％ | -0.16％ |
| SD | 2.64 | 0.03 | 0.16 | 0.02 | 0.05 | 10.18 |
| CV | 3.07％ | 1.23％ | 3.12％ | 2.33％ | 3.46％ | 2.69％ |
The calibration curves of this embodiment are shown in fig. 1-6, respectively.
Repetitive data for this example:
the data for the concentration (80 mg/L) are shown in Table 2:
TABLE 2
| Repeatability of | Urea oxalic acid | Uric acid | Urine citric acid | Urinary calcium | Urine phosphorus | Urine magnesium |
| 1 | 82.32 | 1249.2 | 12.73 | 4.34 | 6.68 | 2.8 |
| 2 | 81.55 | 1244.6 | 13.02 | 4.29 | 6.72 | 2.83 |
| 3 | 79.66 | 1248.1 | 12.94 | 4.4 | 6.63 | 2.76 |
| 4 | 82.23 | 1361.9 | 13.43 | 4.27 | 6.74 | 2.7 |
| 5 | 80.19 | 1351.8 | 13.23 | 4.31 | 6.66 | 2.6 |
| 6 | 79.33 | 1351 | 13.35 | 4.2 | 6.58 | 2.7 |
| 7 | 78.98 | 1388.7 | 13.25 | 4.32 | 6.27 | 2.67 |
| 8 | 76.89 | 1379.9 | 13.28 | 4.19 | 6.34 | 2.71 |
| 9 | 79.08 | 1366.5 | 13.21 | 4.19 | 6.26 | 2.7 |
| 10 | 80.23 | 1265.9 | 13.92 | 4.2 | 6.37 | 2.69 |
| 11 | 81.90 | 1283.9 | 13.82 | 4.18 | 6.44 | 2.65 |
| 12 | 82.22 | 1296.2 | 13.79 | 4.3 | 6.42 | 2.68 |
| 13 | 81.45 | 1306.6 | 13.47 | 4.17 | 6.36 | 2.59 |
| 14 | 82.11 | 1272.4 | 14.69 | 4.37 | 6.48 | 2.6 |
| 15 | 84.44 | 1266.2 | 14.2 | 4.28 | 6.46 | 2.5 |
| AVG | 80.84 | 1308.86 | 13.49 | 4.27 | 6.49 | 2.68 |
| SD | 1.88 | 52.33 | 0.51 | 0.07 | 0.16 | 0.08 |
| CV | 2.32％ | 4.00％ | 3.81％ | 1.75％ | 2.51％ | 3.15％ |
The data for the concentration (10 mg/L) are shown in Table 3:
TABLE 3 Table 3
| Repeatability of | Urea oxalic acid | Uric acid | Urine citric acid | Urinary calcium | Urine phosphorus | Urine magnesium |
| 1 | 10.87 | 119.3 | 1.31 | 0.3 | 0.34 | 0.56 |
| 2 | 9.76 | 117.1 | 1.35 | 0.32 | 0.31 | 0.55 |
| 3 | 9.98 | 126.3 | 1.3 | 0.35 | 0.35 | 0.58 |
| 4 | 8.99 | 134.9 | 1.44 | 0.31 | 0.32 | 0.56 |
| 5 | 10.1 | 138.6 | 1.31 | 0.34 | 0.34 | 0.58 |
| 6 | 9.78 | 144.7 | 1.33 | 0.33 | 0.33 | 0.56 |
| 7 | 9.45 | 133 | 1.46 | 0.32 | 0.32 | 0.54 |
| 8 | 8.97 | 132.6 | 1.49 | 0.34 | 0.33 | 0.59 |
| 9 | 9.76 | 132.1 | 1.35 | 0.32 | 0.35 | 0.55 |
| 10 | 9.67 | 124.8 | 1.4 | 0.33 | 0.35 | 0.55 |
| 11 | 9.24 | 126 | 1.38 | 0.35 | 0.36 | 0.51 |
| 12 | 9.45 | 127.1 | 1.36 | 0.34 | 0.34 | 0.54 |
| 13 | 10.02 | 119.1 | 1.43 | 0.32 | 0.35 | 0.54 |
| 14 | 9.69 | 116.8 | 1.36 | 0.34 | 0.36 | 0.59 |
| 15 | 9.56 | 116.6 | 1.33 | 0.35 | 0.35 | 0.58 |
| AVG | 9.69 | 127.27 | 1.37 | 0.33 | 0.34 | 0.56 |
| SD | 0.47 | 8.62 | 0.06 | 0.02 | 0.02 | 0.02 |
| CV | 4.87％ | 6.77％ | 4.27％ | 4.64％ | 4.45％ | 3.99％ |
Example 2:
this embodiment is based on embodiment 1, and differs from embodiment 1 in that: in the embodiment, each reagent is used for placing the urinary calculus risk factor detection kit into a reagent sample tray of a BA200 instrument, placing the collected specimen into a sample position, automatically detecting by the instrument and automatically calculating the result.
The accuracy data of this example are shown in table 4:
TABLE 4 Table 4
| Accuracy of | Urea oxalic acid | Urinary calcium | Urine citric acid | Urine magnesium | Urine phosphorus | Uric acid |
| Theoretical concentration | 90 | 2.19 | 5.2 | 0.864 | 1.39 | 379 |
| 1 | 87.92 | 2.17 | 5.08 | 0.828 | 1.33 | 386 |
| 2 | 89.39 | 2.18 | 4.89 | 0.868 | 1.31 | 374 |
| 3 | 88.07 | 2.16 | 5.31 | 0.879 | 1.41 | 364 |
| 4 | 90.71 | 2.14 | 5.44 | 0.870 | 1.36 | 385 |
| 5 | 87.14 | 2.17 | 5.29 | 0.877 | 1.31 | 361 |
| 6 | 88.91 | 2.17 | 5.17 | 0.899 | 1.43 | 389 |
| 7 | 90.98 | 2.12 | 5.19 | 0.892 | 1.41 | 376 |
| 8 | 87.82 | 2.21 | 5.02 | 0.894 | 1.42 | 363 |
| 9 | 84.99 | 2.20 | 5.13 | 0.871 | 1.37 | 368 |
| 10 | 85.83 | 2.15 | 5.22 | 0.849 | 1.42 | 385 |
| Average value of | 88.18 | 2.17 | 5.17 | 0.87 | 1.38 | 375.28 |
| Relative deviation of | -2.03％ | -1.11％ | -0.49％ | 1.00％ | -1.07％ | -0.98％ |
| SD | 1.92 | 0.03 | 0.16 | 0.02 | 0.05 | 10.57 |
| CV | 2.18％ | 1.20％ | 3.03％ | 2.46％ | 3.50％ | 2.82％ |
Repetitive data for this example:
the data for the concentration (80 mg/L) are shown in Table 5:
TABLE 5
The data for reproducibility at concentrations (10 mg/L) are shown in Table 6:
TABLE 6
| Repeatability of | Urea oxalic acid | Uric acid | Urine citric acid | Urinary calcium | Urine phosphorus | Urine magnesium |
| 1 | 9.87 | 116.3 | 1.22 | 0.31 | 0.34 | 0.58 |
| 2 | 8.97 | 119.7 | 1.26 | 0.29 | 0.31 | 0.57 |
| 3 | 8.98 | 118.6 | 1.36 | 0.31 | 0.35 | 0.61 |
| 4 | 9.87 | 124.6 | 1.34 | 0.3 | 0.32 | 0.58 |
| 5 | 9.14 | 121.3 | 1.36 | 0.29 | 0.34 | 0.60 |
| 6 | 9.60 | 125.8 | 1.29 | 0.27 | 0.33 | 0.56 |
| 7 | 9.19 | 123.2 | 1.31 | 0.31 | 0.33 | 0.56 |
| 8 | 10.28 | 121.9 | 1.36 | 0.32 | 0.33 | 0.61 |
| 9 | 9.88 | 119.1 | 1.27 | 0.28 | 0.35 | 0.59 |
| 10 | 8.95 | 121.2 | 1.23 | 0.33 | 0.35 | 0.57 |
| 11 | 9.74 | 120 | 1.26 | 0.31 | 0.36 | 0.53 |
| 12 | 9.95 | 123.5 | 1.16 | 0.29 | 0.34 | 0.54 |
| 13 | 10.12 | 115.7 | 1.33 | 0.31 | 0.35 | 0.56 |
| 14 | 9.99 | 113.7 | 1.29 | 0.31 | 0.36 | 0.61 |
| 15 | 9.86 | 116.9 | 1.38 | 0.29 | 0.37 | 0.62 |
| AVG | 9.63 | 120.10 | 1.29 | 0.30 | 0.34 | 0.58 |
| SD | 0.45 | 3.47 | 0.06 | 0.02 | 0.02 | 0.03 |
| CV | 4.72％ | 2.89％ | 4.82％ | 5.30％ | 4.80％ | 4.82％ |
The foregoing description of the embodiments has been provided for the purpose of illustrating the general principles of the invention, and is not meant to limit the scope of the invention, but to limit the invention to the particular embodiments, and any modifications, equivalents, improvements, etc. that fall within the spirit and principles of the invention are intended to be included within the scope of the invention.
Claims (6)
1. The urinary system Dan Chengdan risk factor detection and test system is characterized by comprising a matched instrument and a kit, wherein the kit is internally provided with a plurality of reagents, and the plurality of reagents comprise a urinary oxalic acid detection reagent, a urinary calcium detection reagent, a urinary citric acid detection reagent, a urinary magnesium detection reagent, a urinary phosphorus detection reagent and a urinary uric acid detection reagent;
the uric acid detection reagent comprises a first R1 reagent and a first R2 reagent;
the first R1 reagent comprises the following components:
citric acid, DMAB and MBTH, wherein the concentration of the citric acid in the first R1 reagent is 0.01-1M, and the concentration of the DMAB in the first R1 reagent is 0.1-0.5mM; the concentration of MBTH in the first R1 reagent is 0.01-0.5mM;
the first R2 reagent comprises the following components:
buffer solution with pH value of 3.6-6, oxalic acid oxidase and peroxidase; the concentration of the buffer solution in the first R2 reagent is 0.01-0.5M, the concentration of the oxalic acid oxidase in the first R2 reagent is more than or equal to 50U/L, and the concentration of the peroxidase in the first R2 reagent is more than or equal to 5000U/L;
the first R2 reagent is in a dry powder state, and is re-dissolved by water when in use;
the urine citric acid detection reagent comprises a second R1 reagent and a second R2 reagent;
the second R1 reagent comprises the following components: tris buffer solution, NADH and sodium azide, wherein the concentration of the Tris buffer solution is 40mmol/L, the concentration of NADH is 0.4mmol/L, and the concentration of sodium azide is 0.95g/L;
the second R2 reagent includes an R2a reagent and an R2b reagent:
the R2a reagent comprises PIPES and sodium azide, the PIPES concentration is 600mmol/L, and the sodium azide concentration is 0.95g/L;
the R2b reagent comprises malate dehydrogenase and citrate lyase, and the concentration of the malate dehydrogenase is more than 40KU/L; the concentration of the citrate lyase is more than 1KU/L;
the R2b reagent is in a dry powder state, and R2a and R2b are mixed to obtain a reagent second R2 reagent when in use;
the urine calcium detection reagent comprises an R reagent, wherein the R reagent comprises the following components;
azo arsenic III and imidazole; the concentration of azo arsenic III is 0.2mmol/L, and the concentration of imidazole is 75mmol/L;
the magnesium urine detection reagent comprises a third R1 reagent and a third R2 reagent:
the third R1 reagent comprises the following components:
carbonate buffer and glycol ether diamine tetraacetic acid; the concentration of the carbonic acid buffer solution is 0.1mol/L, and the concentration of the glycol ether diamine tetraacetic acid is 0.1mmol/L;
the third R2 reagent comprises dimethylaniline blue and glycine; the concentration of the dimethylaniline blue is 0.5mmol/L; glycine concentration was 25mmol/L;
the urine phosphorus detection reagent comprises a fourth R1 reagent and a fourth R2 reagent;
the fourth R1 reagent comprises sodium chloride with the concentration of 154mmol/L;
the fourth R2 reagent comprises sodium chloride and ammonium molybdate; the concentration of sodium chloride is 154mmol/L; the concentration of ammonium molybdate is 3.5mmol/L;
the urine acid detection reagent comprises the following components:
phosphate, uricase, peroxidase, 4-aminoantipyrine and ascorbate oxidase, wherein the phosphate concentration is 100mmol/L; uricase concentration is greater than 0.12U/ml; the concentration of peroxidase is more than or equal to 1U/mL;4 aminoantipyrine concentration was 0.5mmol/L; the concentration of ascorbate oxidase is greater than 5U/mL.
2. The urinary tract knot Dan Chengdan risk factor detection test system of claim 1, wherein the first R1 reagent is prepared as follows:
dissolving DMAB in methanol to prepare DMAB mother liquor, dissolving MBTH in water to prepare MBTH mother liquor, dissolving citric acid in water to prepare citric acid solution, and mixing the DMAB mother liquor, the MBTH mother liquor and the citric acid solution in proportion to obtain the first R1 reagent.
3. The urinary tract knot Dan Chengdan risk factor detection test system of claim 1, wherein the first R2 reagent is prepared as follows:
the oxalic acid oxidase is redissolved with water to form a solution and stored at 2-8 ℃, the peroxidase is redissolved with water to form a solution and stored at 2-8 ℃, and the buffer solution, the oxalic acid oxidase aqueous solution and the peroxidase aqueous solution are mixed according to a proportion to obtain a first R2 reagent.
4. The urinary tract knot Dan Chengdan risk factor detection test system of claim 1, wherein the second R2 reagent is prepared as follows:
PIPES and sodium azide are mixed according to a proportion to form R2a, malate dehydrogenase is redissolved to form a solution, citrate lyase is redissolved to form a solution, and the two enzymes are mixed according to a proportion to form R2b and are freeze-dried to form dry powder.
5. The urinary system Dan Chengdan risk factor detection test system of claim 1, wherein the oxalic acid detection reagent, the calcium urinary detection reagent, the citric acid urinary detection reagent, the magnesium urinary detection reagent, the phosphorus urinary detection reagent, and the uric acid detection reagent are arranged in zones.
6. The urinary tract knot Dan Chengdan risk factor detection test system of any one of claims 1-5, wherein the instrument comprises a BST300 and a BA200.
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| US4921807A (en) * | 1985-10-15 | 1990-05-01 | Mission Pharmacal Company | Method and apparatus for maintaining urine specimens |
| CN104198473A (en) * | 2014-08-14 | 2014-12-10 | 上海睿康生物科技有限公司 | Stable uric acid detection kit |
| CN104677959A (en) * | 2015-03-17 | 2015-06-03 | 武汉康复得生物科技股份有限公司 | Reagent, kit and test paper for determining concentration of oxalic acid, and preparation method for test paper |
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| US4921807A (en) * | 1985-10-15 | 1990-05-01 | Mission Pharmacal Company | Method and apparatus for maintaining urine specimens |
| CN104198473A (en) * | 2014-08-14 | 2014-12-10 | 上海睿康生物科技有限公司 | Stable uric acid detection kit |
| CN104677959A (en) * | 2015-03-17 | 2015-06-03 | 武汉康复得生物科技股份有限公司 | Reagent, kit and test paper for determining concentration of oxalic acid, and preparation method for test paper |
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