CN108486090A - A kind of serum inorganic phosphorus that stability is strong (enzyme process) detection kit - Google Patents
A kind of serum inorganic phosphorus that stability is strong (enzyme process) detection kit Download PDFInfo
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- CN108486090A CN108486090A CN201810164809.6A CN201810164809A CN108486090A CN 108486090 A CN108486090 A CN 108486090A CN 201810164809 A CN201810164809 A CN 201810164809A CN 108486090 A CN108486090 A CN 108486090A
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Abstract
The present invention relates to clinical vitro detection reagent technique field, more particularly to a kind of serum inorganic phosphorus that stability is strong (enzyme process) detection reagent.Contain Tris pH of buffer 7.5, inosine, purine nucleoside phosphorylase PNP, xanthine oxidase, peroxidase, glycerine, propylene glycol monomethyl ether, 4 methylphenylboronic acids (4 FPBA), D trehaloses, lithium chloride in reagent R1, contain Tris pH of buffer 7.5,2 in reagent R2,4,6 tribromo, 3 hydroxybenzoic acid (TBHBA), NaN3 0.5g/L, the reagent are suitble to match with various automatic clinical chemistry analyzers, and accuracy and anti-interference ability are good, and have good thermal stability.
Description
Technical field
The present invention relates to clinical vitro detection reagent technique field, more particularly to a kind of serum inorganic phosphorus that stability is strong
（Enzyme process）Detection reagent.
Background technology
1. background
(Phosphorus) about 17mol (530g) phosphorous in human body, wherein 87% is present in bone, remaining be present in soft tissue,
Into the cell.For example certain protein of many important substances, ester type compound, nucleic acid, coenzyme etc. all contain phosphorus in vivo, flat in soda acid
Phosphate also plays an important roll in weighing apparatus.
Phosphorus in blood can be divided into 4 parts：Phos, organophosphor or phosphate, containing phospholipid and other a small amount of phosphatizations
Object.Phosphorus in phosphorus and bone in blood maintains dynamic equilibrium, internal phosphorus mainly to be participated in the form of phosphate many to life
The very important substance metabolism process of activity.Such as：Phosphatide is constituted, and then constitutes the constituent of cell with protein；It participates in
Energetic supersession, especially oxidative phosphorylation process form ATP etc.；Participate in DNA, RNA and many coenzyme (such as phosphorylated thiamines
Element, Coenzyme I and Coenzyme I I etc.) composition；Phosphorus participates in a variety of enzymatic reactions also as the activator or inhibitor of certain enzymes, sugar,
Many biochemical reactions during lipid and protein metabolism, are also required for phosphorus to participate in.
2. clinical meaning
Serum inorganic phosphorus increases：Hypoparathyroidism, pseudohypoparathyroidism, hypervitaminosis D, kidney
Insufficiency or failure, the myeloma that mostly occurs and union phase, acute acid poison, leukaemia, lymthoma, bone tumour, old age
Cataract, Pregnancy Induced Hypertension.Hyperphosphatemia is often asymptomatic, but when calcium, phosphorus concentration (mg/dl) product are more than 70, can occur
Metastatic calcification causes calcium salt to think that precipitation, clinic are closed in joint, ligament, cardiac muscle, blood vessel, subcutaneous, cornea, conjunctiva etc.
Save the symptoms such as pain, eye conjunctivitis, keratitis and pruitus.In addition, hyperphosphatemia be often accompanied by different degrees of hypocalcemia and
The related symptom of renal failure.
Serum inorganic phosphorus reduces：Hyperparathyroidism, rickets or osteomalacia, sugar are become using increase, renal tubule
Venereal disease change, chylous diarrhea, hyperthyroidism, hepatic sclerosis and pulmonary heart disease.
3. detection method
The method that determination of inorganic phosphorus uses at present has colorimetric method, spectrophotometry and enzyme process.
(1) ferrous sulfate occluded corrosion cell
Principle：With trichloroacetic acid protein precipitation, molybdic acid ammonium reagent is added in nonprotein filtrate, is combined with Phos and generates phosphorus molybdenum
Acid, then using ferrous sulfate as reducing agent, it is reduced into blue compound, carry out colorimetric estimation.
Evaluation：This method needs sample removing protein, and complex for operation step, error is big.
(2) ultraviolet spectrophotometry
Principle：Phos is formed by compound with ammonium molybdate effect in an acidic solution in serum, directly in 340nm or
325nm wavelength measures its absorbance.
Evaluation：This method color stability, reagent configuration is simple, and is suitable for automatic clinical chemistry analyzer, but due to serum
In there are reducing substances, be easy to cause testing result inaccuracy.
(3) xanthine oxidase colorimetric method
Principle：Phosphate anion under the catalysis of purine nucleoside phosphorylase (PNP), generates time Huang with inosinic acid
Purine and ribose -1- phosphoric acid.Xanthine oxidase aoxidizes hypoxanthine and generates uric acid and hydrogen peroxide.It is in Trinder reactions
Color, the variation of absorbance are proportional to the concentration of phosphate anion.
Evaluation：This method is easy to operate, has good linear, repeatability and anti-interference ability, is suitble to full-automatic biochemical
Analyzer.
Invention content
It is directed to serum inorganic phosphorus detection, the present invention provides a kind of detections of serum inorganic phosphorus that stability is strong (enzyme process) to try
Agent box, the kit have good thermal stability, and using simply, conveniently, have good compared with common detection methods
Accuracy, repeatability and good detection range are conducive to the popularization and application of reagent clinically.
What the present invention was obtained through the following steps：
A kind of serum inorganic phosphorus that stability is strong (enzyme process) detection kit, including reagent R1, R2, constituent are as follows：
R1：
Tris buffer solutions（pH7.5） 50mmol/L
Inosine 0.3mmol/L
Purine nucleoside phosphorylase PNP 100U/L
Xanthine oxidase 20U/L
Peroxidase 2KUL
Glycerine 1%
Propylene glycol-monomethyl ether 20mmol/L
4- methylphenylboronic acids (4-FPBA) 0.01%
D- trehaloses 5g/L
Lithium chloride 5mmol/L
R2：
Tris buffer solutions（pH7.5） 50mmol/L
Tri- bromo- 3- hydroxybenzoic acids (TBHBA) 5mmol/L of 2,4,6-
NaN3 0.5g/L。
A kind of serum inorganic phosphorus that stability is strong (enzyme process) detection kit, which is characterized in that the reagent is when in use
R1：R2= 5:1, R1 dosage is 250 μ l.
The beneficial effects of the invention are as follows：
（1）Present invention uses lithium chloride, glycerine, D- trehaloses, propylene glycol-monomethyl ether and 4- methylphenylboronic acids to stablize as enzyme
Agent ensure that the enzyme in reagent can stablize 12d or more under the conditions of 37 DEG C.
（2）Present invention uses Tris hydrochloride buffers ensure that by the pH stable of reagent in the optimum level of enzyme
The stability of reagent improves the catalytic efficiency of enzyme.
Description of the drawings
Fig. 1 is 1 reagent test method of embodiment；
Fig. 2 is 1 reagent interference free performance comparison result of embodiment；
Fig. 3 is 1 reagent of embodiment and control group kit contrasting detection result；
Fig. 4 is 1 reagent of embodiment and control group kit correlation curve figure；
Fig. 5 be 1 reagent of embodiment with compare group reagent thermal stability curve graph.
Specific implementation mode
With reference to specific embodiment, invention is further explained：
(1) embodiment 1
R1：
Tris buffer solutions（pH7.5） 50mmol/L
Inosine 0.3mmol/L
Purine nucleoside phosphorylase PNP 100U/L
Xanthine oxidase 20U/L
Peroxidase 2KUL
Glycerine 1%
Propylene glycol-monomethyl ether 20mmol/L
4- methylphenylboronic acids (4-FPBA) 0.01%
D- trehaloses 5g/L
Lithium chloride 5mmol/L
R2：
Tris buffer solutions（pH7.5） 50mmol/L
Tri- bromo- 3- hydroxybenzoic acids (TBHBA) 5mmol/L of 2,4,6-
NaN3 0.5g/L。
The application method of the present embodiment reagent：
The present embodiment reagent uses automatic clinical chemistry analyzer, such as 7180 fully-automatic analyzer of Hitachi when in use, utilizes speed
Rate method is measured.The ratio of sample and reagent is set as 10：250：50, Detection wavelength 505nm, in the correspondence of sample disc
Position places distilled water, standard items and sample, and Fig. 1 is shown in operation.
Embodiment 2
Interference is tested：Fresh mix serum is taken, 2 equal portions are divided into, then will be separated into 5 equal portions per equal portions, different do is added
Substance is disturbed, its concentration in serum is made to reach the requirement of Fig. 2.Then 1 gained reagent of embodiment is used respectively, it is common simultaneously with market
The serum inorganic phosphorus reagent of approval while the content of comparative determination serum inorganic phosphorus, control group measurement result and addition disturbance
The measurement result of each group is shown in Fig. 2 after substance.Relative deviation (%)=(measurement of measurement mean value-check sample of interference sample is equal
Value)/check sample measurement mean value × 100%.
As seen from Figure 2,1 reagent of embodiment is in bilirubin≤300 μm ol/L, creatinine≤200 μm ol/L, hemoglobin
Test result is not significantly interfered in the case of≤200mg/L, triglycerides≤2.5mmol/L, cholesterol≤50 μm ol/L.With
Control group result is compared, and reagent anti-interference ability of the present invention is preferable, illustrates the reagent of embodiment 1 in accuracy and anti-interference ability
It is upper up to standard.
Embodiment 3
Correlation is tested：Using 1 reagent of embodiment, certain company approved with the common State Food and Drug Administration in market
Serium inorganic phosphorus (enzyme process) kit as a contrast group reagent carry out control test, while detect 20 clinical serum samples, detection knot
Fruit is as shown in figure 3, and obtain the correlation curves of two kinds of reagents（As shown in Figure 4）.It is shown by testing result, two reagents
The related coefficient of box is 0.999, illustrates that the two has great correlation.
Experiment calibration object used and quality-control product are respectively：
Calibration object：The content of serum inorganic phosphorus is 1.43 mmol/L.
Quality-control product：The target value of serum inorganic phosphorus is 1.41mmol/L, target value range：1.20-1.62mmol/L.
Embodiment 4
Reagent stability verification test：Using 1 reagent of the embodiment of the present invention as test group, a kind of commercially available serum inorganic phosphorus (enzyme is taken
Method) detection kit as a control group, test group asks for portion with every group of group reagent is compareed, and does the examination of 37 DEG C of heat stability tests
It tests, closing is placed in 37 DEG C of thermostat water baths（It is only taken out when detection daily, after detection, still sealing is put back to
In 37 DEG C of water-baths, continuous 12 days）, verified as 37 DEG C of thermal stability.By reagent simultaneously in 7180 full-automatic biochemical of Hitachi point
On analyzer device, it is detected according to Fig. 1 methods, and standard curve is established on instrument.Freeze-dried powder quality-control product is taken, is uniformly dissolved
Afterwards, 15 parts are divided into, -20 DEG C store, daily Quality Control one, and tracing detection is as a result, its thermal stability tracking and monitoring becomes
Gesture such as Fig. 5.
Claims (4)
1. a kind of serum inorganic phosphorus that stability is strong (enzyme process) detection kit, which is characterized in that be made of reagent R1, R2, group
At ingredient：
R1：
Tris buffer solutions（pH7.5） 50mmol/L
Inosine 0.3mmol/L
Purine nucleoside phosphorylase PNP 100U/L
Xanthine oxidase 20U/L
Peroxidase 2KUL
Glycerine 1%
Propylene glycol-monomethyl ether 20mmol/L
4- methylphenylboronic acids (4-FPBA) 0.01%
D- trehaloses 5g/L
Lithium chloride 5mmol/L
R2：
Tris buffer solutions（pH7.5） 50mmol/L
Tri- bromo- 3- hydroxybenzoic acids (TBHBA) 5mmol/L of 2,4,6-
NaN3 0.5g/L。
2. a kind of serum inorganic phosphorus that stability is strong (enzyme process) detection kit, which is characterized in that reagent R1 when in use：
R2= 5:1, R1 dosage is 250 μ l.
Stablize as enzyme 3. present invention uses lithium chloride, glycerine, D- trehaloses, propylene glycol-monomethyl ether and 4- methylphenylboronic acids
Agent ensure that the enzyme in reagent can stablize 12d or more under the conditions of 37 DEG C.
4. present invention uses Tris hydrochloride buffers, by the pH stable of reagent in the optimum level of enzyme, reagent ensure that
Stability improves the catalytic efficiency of enzyme.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810164809.6A CN108486090A (en) | 2018-02-27 | 2018-02-27 | A kind of serum inorganic phosphorus that stability is strong (enzyme process) detection kit |
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| CN201810164809.6A CN108486090A (en) | 2018-02-27 | 2018-02-27 | A kind of serum inorganic phosphorus that stability is strong (enzyme process) detection kit |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108120713A (en) * | 2017-12-20 | 2018-06-05 | 青岛汉唐生物科技有限公司 | A kind of Tes-Tape and preparation method thereof |
| CN110596372A (en) * | 2019-08-09 | 2019-12-20 | 山东博科生物产业有限公司 | Potassium ion detection kit |
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| CN1778954A (en) * | 2004-11-23 | 2006-05-31 | 苏州艾杰生物科技有限公司 | Determination of inorganic phosphorus and its diagnostic reagent kit |
| CN101620141A (en) * | 2008-06-30 | 2010-01-06 | 苏州艾杰生物科技有限公司 | Inorganic phosphorus (phosphate radical) diagnosis/determination kit and method for determining inorganic phosphorus (phosphate radical) concentration |
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2018
- 2018-02-27 CN CN201810164809.6A patent/CN108486090A/en active Pending
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|---|---|---|---|---|
| CN1778954A (en) * | 2004-11-23 | 2006-05-31 | 苏州艾杰生物科技有限公司 | Determination of inorganic phosphorus and its diagnostic reagent kit |
| CN101620141A (en) * | 2008-06-30 | 2010-01-06 | 苏州艾杰生物科技有限公司 | Inorganic phosphorus (phosphate radical) diagnosis/determination kit and method for determining inorganic phosphorus (phosphate radical) concentration |
| CN101955923A (en) * | 2010-09-30 | 2011-01-26 | 江南大学 | Method for improving storage stability of catalase |
| CN103642781A (en) * | 2013-12-12 | 2014-03-19 | 山东博科生物产业有限公司 | Horse radish peroxidase protective agent |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108120713A (en) * | 2017-12-20 | 2018-06-05 | 青岛汉唐生物科技有限公司 | A kind of Tes-Tape and preparation method thereof |
| CN110596372A (en) * | 2019-08-09 | 2019-12-20 | 山东博科生物产业有限公司 | Potassium ion detection kit |
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