CN106885905A - A kind of urea detection reagent and detection method with superior detection line and sensitivity for analysis - Google Patents
A kind of urea detection reagent and detection method with superior detection line and sensitivity for analysis Download PDFInfo
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- CN106885905A CN106885905A CN201510942729.5A CN201510942729A CN106885905A CN 106885905 A CN106885905 A CN 106885905A CN 201510942729 A CN201510942729 A CN 201510942729A CN 106885905 A CN106885905 A CN 106885905A
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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Abstract
The present invention relates to UREA detection technique fields, more particularly to a kind of UREA detection reagents contain TRIS buffer, NADPH, preservative in reagent R1;Contain TRIS buffer, urase, glutamte dehydrogenase, KG, XOD, POD, preservative in reagent R2.Improvement enzyme process urea reagent has more superior test limit and assay sensitivity, and self performance is good, greatly strengthen the stability of reagent, meets relevant clinical application requirement.
Description
Technical field
The present invention relates to urea detection technique field, more particularly to a kind of urine with superior detection line and sensitivity for analysis
Plain detection reagent, further relates to use the detection method of this detection reagent.
Background technology
Urea(UREA)It is the dead end product of human body protein metabolism.Vivo acid resolves into α -one through deamination
Acid and NH3, NH3Enter urea cycle and CO in liver cell2Generation urea.The growing amount of urea is taken the photograph depending on dietary protein
Enter the catabolism and liver function of amount, tissue protein.The urea of generation is discharged through blood circulation is main by kidney.Through saliva
Liquid, gastric juice, bile and intestinal juice drain into the urea in alimentary canal, and the overwhelming majority resolves into NH3Again through liver urea synthesis after absorption
Still from RE.Urea in blood plasma can all from glomerular filtration, and about 30% ~ 40% is inhaled again by glomerulus under normal circumstances
Receive, glomerulus also can excretion urea on a small quantity.Plasma urea concentrations can reflect the filtering function of glomerulus to a certain extent, this
Outward, kidney other factor such as tissue catabolism quickening, hemorrhage of digestive tract, hyperphagia protein etc. can all cause plasma urea concentrations
Raise, thus plasma urea concentrations measure is not also the specific parameters of renal dysfunction.Even so, because urea is by kidney
The main component of the low molecule nitrogenouz wastes of excretion, plasma urea concentrations are to the course of disease of chronic renal disease, disease observation and pre-
After judge meaningful, and Plasma Urea assay method is fairly simple, so it is still current kidney trouble that Plasma Urea is determined
One of main inspection project.
At present, the assay method of the urea activity of document report has urease methods, phenol-hypochlorite development process, Na Shi to try
Agent development process, diacetyl monoxime method etc..These method accuracys are poor, sensitivity is low, instrument and equipment requirement is high, reagent cost is high,
It is difficult to popularize application in an all-round way.
In consideration of it, the present invention is improved to existing enzyme process urea detection reagent, and its performance indications is commented
Estimate.Improvement enzyme process urea reagent has more superior test limit and assay sensitivity, and self performance is good, meets correlation
Clinical practice requirement.
The content of the invention
The side of UREA contents is detected it is an object of the invention to provide a kind of reagent for detecting UREA and using the reagent
Method.The kit uses urase-glutamate dehydrogenase enzyme process, can be with the content of effective detection UREA, strong antijamming capability, stability
The advantages of good.
General principle:Urea generates NH4+ and HCO3- under urase effect,
Urase
Urea+H2O2NH4++2HCO3-
NH4 +And KG, reduced coenzyme Ⅰ are under glutamate dehydrogenase enzymatic, Pidolidone and NAD are generated+
Glutamte dehydrogenase
KG+NH4++ NADHPidolidone+NAD++ H2O
By the rate of change for determining NADH absorbances at 340nm wavelength(ΔA/min)UREA activity can be measured.
What the present invention was obtained through the following steps:
A kind of sour test agent of UREA inspections, including reagent R1 and reagent R2, the composition of the reagent R1 and reagent R2 are as follows:
1)The component of reagent R1 is:
Buffer solution(PH value 9.1)··········································100mmol/L
NADPH····················································0.3mmol/L
2)The component of reagent R2 is:
Buffer solution(PH value 7.6)··········································100mmol/L
Urase·····················································40KU/L
Glutamte dehydrogenase·············································2KU/L
KG···············································30mmol/L
Preservative···················································0.8g/L;
Above reagent uses trishydroxymethylaminomethane cushioning liquid.
Above UREA detection reagents, the preservative is NaN3.
Described UREA detection reagents detect the detection method of UREA, and performance rate method is utilized using automatic clinical chemistry analyzer
It is measured, detection dominant wavelength is 340nm.
The ratio of described detection method, R1 reagents and R2 reagents is 4:1.
Beneficial effects of the present invention:
1)Using new buffer system and stabilizer, the stability of reagent is significantly improved;
2)Improvement enzyme process urea reagent has more superior test limit and assay sensitivity, and self performance is good, meets
Relevant clinical application requirement.
Brief description of the drawings
Fig. 1 is two kinds of correlation curve figures of reagent,
Fig. 2 is two kinds of reagent effect phase stability curve figures.
Specific embodiment
The present invention is further described with reference to specific embodiment:
Embodiment 1
The detection reagent of UREA, bag reagent R1 and reagent R2:
1)The component of reagent R1 is:
Buffer solution(PH value 9.1)···············································100mmol/L
NADPH·························································0.3mmol/L
2)The component of reagent R2 is:
Buffer solution(PH value 7.6)···············································100mmol/L
Urase··························································40KU/L
Glutamte dehydrogenase··················································2KU/L
KG····················································30mmol/L
Preservative························································0.8g/L;
3)The application method of the present embodiment reagent:
The UREA detection reagents of the present embodiment description, when in use using the automatic clinical chemistry analyzer with double reagent function,
Such as the fully-automatic analyzer of Hitachi 7180, is measured using performance rate method.By R1 and R2 according to 4:1 ratio is placed into corresponding
On reagent position, distilled water, standard items and sample, operation such as table 1 are placed in the correspondence position of sample disc:
The reagent test method of 1 embodiment of table 1
Calculate:UREA contents(mmol/L)=(A determines ÷ A standards)× C standards.
Embodiment 2
Interference is tested:Fresh mix serum is taken, is divided into 2 equal portions, 5 equal portions then will be separated into per equal portions, add different doing
Material is disturbed, its concentration in serum is reached the requirement of table 2.Then respectively use the gained reagent of embodiment 1, it is common with market simultaneously
The UREA reagents of accreditation simultaneously in comparative determination serum UREA content, control group measurement result with add disturbance material after
The measurement result of each group is shown in Table 2.Relative deviation(%)=(Disturb the measure average of the measure average-check sample of sample)/ control
Measure average × 100% of sample.
As can be seen from Table 2, the reagent of embodiment 1 is in ascorbic acid≤1704 μm ol/L, bilirubin≤684 μm ol/L, blood
Lactoferrin≤5 g/L, triglycerides≤22.6 mmol/L are not significantly interfered with to test result.And group reagent is compareed above-mentioned
In the presence of concentration interfering material, significantly interfered with, the interference free performance of this explanation reagent of embodiment 1 is far superior to having a competition
Agent.
The embodiment reagent interference free performance of table 2 compares
Embodiment 3
Correlation is tested:Reagent preparation, the State Food and Drug Administration accreditation common with market are formulated using embodiment 1
The UREA kits of certain company carry out control test, while have detected 20 clinical serum samples, the testing result such as institute of table 3
Show.And obtain two kinds of correlation curves of reagent(As shown in Figure 1), shown by testing result, two correlations of kit
Coefficient is 0.9974, illustrates that both have great correlation.
The reagent of 3 embodiment of table 1 and market are common and UREA that get the nod determines kit contrasting detection result
Embodiment 4
The stability contrast test of reagent:To the reagent in embodiment 1, uniform 13 groups of packing, every group of amount of reagent is for R1
40mL, R2 are 10mL;And take 13 groups of UREA examinations of certain company of the common State Food and Drug Administration's accreditation in market
Agent box is compared.It is placed into 2-8 DEG C of refrigerator, the group reagent of taking-up on the same day detection UREA quality-control products monthly, testing result
As shown in Fig. 2 more common than the market UREA under 2-8 DEG C of condition of storage of the reagent of embodiment 1 determines kit more stablizing.
By checking, this reagent is good with similar detection reagent contrast correlation, and clinical detection sample results are consistent, Neng Gouda
To market to the application requirement of product, and good in anti-interference performance, be it is a kind of more stablize, good UREA detection reagents.
Claims (6)
1. a kind of UREA detection reagents, it is characterised in that including reagent R1 and reagent R2, the composition of the reagent R1 and reagent R2
It is as follows:
1)The component of reagent R1 is:
Buffer solution··························································100mmol/L
NADPH···························································0.3mmol/L
2)The component of reagent R2 is:
Buffer solution··························································100mmol/L
Urase····························································40KU/L
Glutamte dehydrogenase····················································2KU/L
KG······················································30mmol/L
Preservative··························································0.8g/L·。
2. UREA detection reagents according to claim 1, it is characterised in that buffer solution is 25 DEG C in reagent R1, and pH is 9.1
TRIS buffer.
3. UREA detection reagents according to claim 1, it is characterised in that buffer solution is 25 DEG C in reagent R2, and pH is 7.6
TRIS buffer.
4. UREA detection reagents according to claim 1, it is characterised in that the preservative is NaN3。
5. the detection method of UREA is detected any one of a kind of usage right requirement 1-4, it is characterised in that using full-automatic
Biochemical Analyzer is measured using performance rate method, and detection dominant wavelength is 340nm.
6. detection method according to claim 5, it is characterised in that the ratio of R1 reagents and R2 reagents is 4:1.
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Cited By (2)
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CN112126674A (en) * | 2020-08-18 | 2020-12-25 | 兰州百源基因技术有限公司 | Urea detection kit, preparation method and use method thereof |
JP2021177167A (en) * | 2020-03-19 | 2021-11-11 | 財團法人工業技術研究院Industrial Technology Research Institute | Detection reagent, detection device, and method for detecting primary amide compound |
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Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
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JP2021177167A (en) * | 2020-03-19 | 2021-11-11 | 財團法人工業技術研究院Industrial Technology Research Institute | Detection reagent, detection device, and method for detecting primary amide compound |
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CN112126674A (en) * | 2020-08-18 | 2020-12-25 | 兰州百源基因技术有限公司 | Urea detection kit, preparation method and use method thereof |
CN112126674B (en) * | 2020-08-18 | 2024-02-06 | 兰州百源基因技术有限公司 | Urea detection kit, preparation method and use method thereof |
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