CN102417921A - Determination reagent against bilirubin interference based on uricase method and application method thereof - Google Patents
Determination reagent against bilirubin interference based on uricase method and application method thereof Download PDFInfo
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- CN102417921A CN102417921A CN2010105011663A CN201010501166A CN102417921A CN 102417921 A CN102417921 A CN 102417921A CN 2010105011663 A CN2010105011663 A CN 2010105011663A CN 201010501166 A CN201010501166 A CN 201010501166A CN 102417921 A CN102417921 A CN 102417921A
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Abstract
The invention relates to a determination reagent against bilirubin interference based on a uricase method and an application method thereof. The reagent is a liquid double-reagent and has good resistance against bilirubin interference and excellent stability. The reagent can be used in a semi/full-automatic biochemical analyzer by adopting a two-point end assay method. By adopting a specific enzyme, the reagent has the characteristics of high sensitivity and strong anti-interference capability; and by adding a selected stabilizing agent, the reagent achieves good stability, namely, the reagent can be hermetically preserved at the temperature of 2-8 DGE C in dark place for 12 months without causing performance deterioration. The reagent can be widely applied to diagnosis, of kidney diseases such as acute and chronic nephritis, kidney stone, gout and the like as well as diseases resulting from excessive nucleic acid metabolism, such as leukemia, multiple myeloma and the like, physical examination and general investigation of kidney diseases and the like.
Description
Technical field
The invention belongs to biological technical field, relate to a kind of anti-UCB interferential uric acid enzymic measuring reagent and method of use thereof.
Background technology
Uric acid (UA) is the final product of purine metabolism in the nucleic acid, and in human body, purine nucleotides through hydrolytic deaminization and oxidation, generates UA after decomposing generation purine nucleoside and purine at last.UA discharges with urine, and UA all leaches through renal glomerulus in the blood, is almost completely heavily absorbed at uriniferous tubules, so the clearance rate of UA is extremely low.The UA that is discharged by kidney accounts for the 2/3-3/4 of total output on the one, and all the other are decomposed by the enzyme of mikrobe at gi tract.UA can not normally drain when GFR lowered, and UA concentration raises in the blood.Some medicines also influence the drainage of UA, can promote the discharge of UA like this semi-annular jade pendant of thiazide diuretic box hydroxyl amine.The Whitfield's ointment preparation also increases its drainage when high dosage.Usually the increase of UA level and nitrogenous substances, urea, creatinine are relevant with other non-albumen factors, are usually used in the not auxiliary diagnosis of the hyperuricemia of bright reason of gout, renal tubal dysfunction, bone marrow disease and some clinically.
Clinical meaning:
(1) content of the excretory function of the accretion rate of UA content and nucleic acid in vivo in blood, kidney and food amplifying nucleic acid is relevant.UA is difficult to dissolving, and concentration Gao Shihui is deposited in the tissue, forms calculus.
(2) serum UA increases and is mainly seen in gout, is the leading indicator of clinical diagnosis gout.Due to the imbalance of uratoma purine metabolism, serum UA can obviously raise.
(3) the hyperfunction endogenous UA that causes of nucleic acid metabolism generates increase, and serum UA rises.Be shown in white blood disease, multiple myeloma, polycythemia vera etc.
(4) gestational hypertension, the pathology that renal blood flows such as eclampsia reduce raises serum UA because of UA drains to reduce.But the normal no change of SUR this moment.
(5) serum UA raises and also is shown in chronic lead poisoning, chloroform and carbon tetrachloride poisoning.
(6) the many because metabolism disorder of Hypouricemia generates not enough, uriniferous tubules like UA and the transhipment of UA is reached in the urine UA unusually discharges due to unusual the increasing, and also is shown in WilsonShi disease, Fancoi syndromes, serious anaemia etc.
Hyperuricemia has caused showing great attention to of medical circle as the sick new kind of modern civilization, detects UA exactly and in clinical application, more seems important.The UA measuring method is a lot, and phospho-wolframic acid method, uric acid enzyme process, dry chemistry method and HPLC method etc. are arranged.Present most popular method is uriKoxidase-peroxidase reaction system, method sensitive and do not need deproteinize, main obscurant be xitix (vitamins C, VC) and UCB.Therefore, solve the interference problem that xitix and UCB are measured UA, significant.
Summary of the invention
Technical problem to be solved by this invention provides a kind of immunity from interference good anti-UCB interferential uric acid enzymic measuring reagent and method of use thereof.
The present invention solves the problems of the technologies described above the technical scheme that is adopted: a kind of anti-UCB interferential uric acid enzymic measuring reagent, and said reagent is liquid double reagent, wherein a kind of component of reagent 1 and content are:
Phosphate buffered saline buffer (pH 6.0-8.0) 0.10-100g/L,
Px 0.10-10KU/L,
N-ethyl-N-(3-toluene) N-succinyl-quadrol EMSE 0.10-10g/L,
Anti-interference enzyme 0.10-10KU/L,
Stablizer 0.10-100g/L;
Wherein the component of second kind of reagent 2 and content are:
Phosphate buffered saline buffer (pH 6.0-8.0) 0.10-100g/L,
Px 0.10-10KU/L,
UriKoxidase 0.10-10KU/L,
4 aminoantipyrene 4-AAP 0.10-10g/L,
Stablizer 0.10-100g/L
Specifically, said anti-interference enzyme is anti-BOX; Said stablizer is at least a in bovine serum albumin, yellow prussiate, Triton-100, polyvalent alcohol, the polyoxyethylene glycol; Wherein phosphate buffered saline buffer can adopt potassium hydrogenphosphate, potassium primary phosphate etc.
A kind of method of use of above-mentioned anti-UCB interferential uric acid enzymic measuring reagent comprises the steps:
(1) sample is prepared: sample is not haemolysis serum or a blood plasma of empty stomach, and sample transports preservation under coldcondition;
(2) adopt automatic clinical chemistry analyzer that the sample of step (1) is detected, concrete parameter is following:
Reaction type: 2 end-point methods,
The Direction of Reaction: positive reaction,
Predominant wavelength: 600nm,
Commplementary wave length: 800nm,
Temperature of reaction: 37 ℃,
Sample/reagent (S/R): 3/200,
Sample size (S): 3 μ l,
Reagent 1 (Ri): 150 μ l,
Reagent 2 (R2): 50 μ l,
Reaction times: 10min.
Insoluble blood serum of empty stomach or blood plasma adopt anticoagulant heparin in the step (1), but 0.1mg heparin anti-freezing 1.0ml blood; Or employing EDTA anti-freezing: but 1.8mg EDTA anti-freezing 1.0ml blood.Uric acid can be stablized 8 hours at ambient temperature in the sample, can stablize 3 days at 2 ℃～8 ℃, can stablize 6 months-20 ℃ of preservations.
In sum, the invention has the beneficial effects as follows: reagent of the present invention is liquid double reagent, has good anti-UCB jamming performance and advantages of excellent stability.Adopt 2 end-point methods, can on half/automatic clinical chemistry analyzer, use.These article have adopted a kind of specific enzyme, make reagent have characteristics highly sensitive, strong anti-interference performance; These article add selected stablizer, make reagent have satisfactory stability property, and preserving (lucifuge sealing) down at 2-8 ℃ did not influence its performance in 12 months.This reagent can be widely used in kidney diseases such as acute and chronic nephritis, urinary stone disease, gout, and the generaI investigation of the diagnosis of containing disease, health examination, kidney disease etc. is crossed in nucleic acid metabolisms such as white blood disease, multiple myeloma.
Embodiment
Embodiment 1:
(1) sample is prepared: sample is not haemolysis serum or blood plasma (anticoagulant heparin, but 0.1mg heparin anti-freezing 1.0ml blood of empty stomach; The EDTA anti-freezing: but 1.8mg EDTA anti-freezing 1.0ml blood).Sample should transport preservation under coldcondition, uric acid can be stablized 8 hours at ambient temperature in the sample, can stablize 3 days at 2 ℃～8 ℃, can stablize 6 months-20 ℃ of preservations.
(2) by following composition and consumption preparation testing uric acid reagent:
Reagent 1:
Phosphate buffered saline buffer (pH 6.0-8.0) 0.10-100g/L,
Px 0.10-10KU/L,
N-ethyl-N-(3-toluene)-N-succinyl-quadrol EMSE 0.10-10g/L,
Anti-interference enzyme 0.10-10KU/L,
Stablizer 0.10-100g/L;
Reagent 2:
Phosphate buffered saline buffer (pH 6.0-8.0) 0.10-100g/L,
Px 0.10-10KU/L,
UriKoxidase 0.10-10KU/L,
4-aminoantipyrene 4-AAP 0.10-10g/L,
Stablizer 0.10-100g/L.
Above-mentioned each component and content thereof are specially; Phosphate buffered saline buffer is the arbitrary value between the 0.10-100g/L in the reagent 1; Be specially 0.1,0.2,0.5,1,3,5,10,20,30,40,50,60,70,80,90,100g/L all can; PH is the arbitrary value between the 6.0-8.0, is specially 6,6.2,6.5,6.7,7.0,7.2,7.5,7.7,8.0 and all can; Px is the arbitrary value between the 0.10-10KU/L, is specially 0.1,0.2,0.5,0.8,1,3,5,7,10KU/L all can; N-ethyl-N-(3-toluene)-N-succinyl-quadrol EMSE is the arbitrary value between the 0.10-10g/L, is specially 0.1,0.3,0.5,1,3,5,10g/L all can; Anti-interference enzyme is the arbitrary value between the 0.10-10KU/L, is specially 0.1,0.2,0.5,0.8,1,3,5,7,10KU/L all can; Stablizer is the arbitrary value between the 0.10-100g/L, is specially 0.1,0.2,0.5,1,3,5,10,20,30,40,50,60,70,80,90,100g/L all can.Phosphate buffered saline buffer is the arbitrary value between the 0.10-100g/L in the reagent 2; Be specially 0.1,0.2,0.5,1,3,5,10,20,30,40,50,60,70,80,90,100g/L all can; PH is the arbitrary value between the 6.0-8.0, is specially 6,6.2,6.5,6.7,7.0,7.2,7.5,7.7,8.0 and all can; Px is the arbitrary value between the 0.10-10KU/L, is specially 0.1,0.2,0.5,0.8,1,3,5,7,10KU/L all can; UriKoxidase is the arbitrary value between the 0.10-10KU/L, is specially 0.1,0.2,0.5,0.8,1,3,5,7,10KU/L all can; The 4-aminoantipyrene is the arbitrary value between the 0.10-10g/L, is specially 0.1,0.3,0.5,1,3,5,10g/L all can; Stablizer is the arbitrary value between the 0.10-100g/L, is specially 0.1,0.2,0.5,1,3,5,10,20,30,40,50,60,70,80,90,100g/L all can.
After the mentioned reagent dissolving is accomplished, divide the bottle of packing into, process liquid double reagent, directly use.On the TBA-120FR of Toshiba type automatic clinical chemistry analyzer according to following parameter setting:
Reaction type: 2 end-point methods,
The Direction of Reaction: positive reaction,
Predominant wavelength: 600nm,
Commplementary wave length: 800nm,
Temperature of reaction: 37 ℃,
Sample/reagent (S/R): 3/200,
Sample size (S): 3 μ l,
Reagent 1 (R1): 150 μ l,
Reagent 2 (R2): 50 μ l,
Reaction times: 10min.
During mensuration, sample adds reagent 1 and is incubated 300 seconds down at 37 ℃, the record absorbance A
1, behind the adding reagent 2, be incubated 300 seconds down, the record absorbance A at 37 ℃
2, calculate the sample uric acid concentration by following formula:
Below through the related experiment data to anti-UCB interferential uric acid enzymic measuring reagent of the present invention, its beneficial effect is further specified, concrete experimental data is following:
1, anti-triglyceride disturbs (mg/dL)
(1) prior art single reagent (A reagent)
(2) prior art double reagent (B reagent)
(3) reagent of the present invention (C reagent)
2, anti-hemolysis disturbs (mg/dL)
(1) prior art single reagent (A reagent)
(2) prior art double reagent (B reagent)
(3) reagent of the present invention (C reagent)
3, anti-UCB disturbs (mg/dL)
(1) prior art single reagent (A reagent)
(2) prior art double reagent (B reagent)
(3) reagent of the present invention (C reagent)
5, uncork stability
Level one quality control product (low value): lot number, 567UN; Target value 340 μ mol/L, scope 300-390 μ mol/L;
Level two quality control products (high value): lot number, 413UE; Target value 560 μ mol/L, scope 490-630 μ mol/L.
Unit: mol/L
Conclusion: uncork 30 days is measured result's the relative error of result and the 1st day all less than 10%, meets the requirement of standard.The component and the content of prior art single reagent (A reagent) are:
Phosphate buffered saline buffer (pH 7.5) 100mmol/L,
Px 10KU/L,
DHBS 2.0mmol/L，
4-aminoantipyrene 4-AAP 4.0mmol/L,
UriKoxidase 150U/L.
The component and the content of prior art double reagent (B reagent) are:
Reagent 1:
Phosphate buffered saline buffer (pH 7.0) 100mmol/L,
TBHBA 1mmol/L；
Reagent 2:
Phosphate buffered saline buffer (pH 7.0) 100mmol/L,
4-aminoantipyrene 4-AAP 0.3mmol/L,
Px 2KU/L,
UriKoxidase 3U/L.
Experiment showed, that through above-mentioned reagent interference free performance of the present invention obviously is superior to the single reagent and the double reagent of prior art, has good anti-bilirubinic interference effect.
Claims (5)
1. an anti-UCB interferential uric acid enzymic measuring reagent is characterized in that said reagent is liquid double reagent, and wherein the component of first kind of reagent 1 and content are:
Phosphate buffered saline buffer (pH 6.0-8.0) 0.10-100g/L,
Px 0.10-10KU/L,
N-ethyl-N-(3-toluene)-N-succinyl-quadrol EMSE 0.10-10g/L,
Anti-interference enzyme 0.10-10KU/L,
Stablizer 0.10-100g/L;
Wherein the component of second kind of reagent 2 and content are:
Phosphate buffered saline buffer (pH 6.0-8.0) 0.10-100g/L,
Px 0.10-10KU/L,
UriKoxidase 0.10-10KU/L,
4-aminoantipyrene 4-AAP 0.10-10g/L,
Stablizer 0.10-100g/L.
2. anti-UCB interferential uric acid enzymic measuring reagent according to claim 1 is characterized in that said anti-interference enzyme is anti-BOX, and content is 0.1-10KU/L.
3. anti-UCB interferential uric acid enzymic measuring reagent according to claim 1 is characterized in that, said stablizer is at least a in bovine serum albumin, yellow prussiate, Triton-100, polyvalent alcohol, the polyoxyethylene glycol.
4. the method for use of the described anti-UCB interferential uric acid enzymic measuring reagent of claim 1 is characterized in that, comprises the steps:
(1) sample is prepared: sample is not haemolysis serum or a blood plasma of empty stomach, and sample transports preservation under coldcondition;
(2) adopt automatic clinical chemistry analyzer that the sample of step (1) is detected, concrete parameter is following:
Reaction type: 2 end-point methods,
The Direction of Reaction: positive reaction,
Predominant wavelength: 600nm,
Commplementary wave length: 800nm,
Temperature of reaction: 37 ℃,
Sample/reagent (S/R): 3/200,
Sample size (S): 3 μ 1,
Reagent 1 (R1): 150 μ l,
Reagent 2 (R2): 50 μ l,
Reaction times: 10min.
5. according to the method for use of the said anti-UCB interferential uric acid enzymic measuring reagent of claim 4, it is characterized in that said empty stomach is haemolysis serum or blood plasma employing anticoagulant heparin not, but 0.1mg heparin anti-freezing 1.0ml blood; Or employing EDTA anti-freezing: but 1.8mg EDTA anti-freezing 1.0ml blood.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010105011663A CN102417921A (en) | 2010-09-25 | 2010-09-25 | Determination reagent against bilirubin interference based on uricase method and application method thereof |
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| CN2010105011663A CN102417921A (en) | 2010-09-25 | 2010-09-25 | Determination reagent against bilirubin interference based on uricase method and application method thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103278648A (en) * | 2013-05-24 | 2013-09-04 | 宁波美康生物科技股份有限公司 | Albumin detection reagent |
| 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 |
| CN104198686A (en) * | 2014-08-14 | 2014-12-10 | 苏州康铭诚业医用科技有限公司 | Compound stabilizer for uric acid assay kit |
| CN104198686B (en) * | 2014-08-14 | 2017-01-04 | 苏州康铭诚业医用科技有限公司 | A kind of testing uric acid test kit |
| CN106404758A (en) * | 2015-07-28 | 2017-02-15 | 珠海和凡医药股份有限公司 | Test paper for detecting uric acid content range in urine |
| CN109916891A (en) * | 2019-04-12 | 2019-06-21 | 吉林省汇酉生物技术股份有限公司 | A kind of dry chemistry reagent piece and preparation method thereof quantitative determining uric acid concentration |
| CN114200122A (en) * | 2021-11-09 | 2022-03-18 | 深圳市锦瑞生物科技股份有限公司 | Freeze-dried reagent ball for uric acid detection, configuration method thereof and microfluidic detection chip |
-
2010
- 2010-09-25 CN CN2010105011663A patent/CN102417921A/en active Pending
Non-Patent Citations (2)
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| 王缚鲲等: "各种胆红素对氧化酶法测定血清尿酸浓度的影响", 《检验医学》 * |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103278648A (en) * | 2013-05-24 | 2013-09-04 | 宁波美康生物科技股份有限公司 | Albumin detection reagent |
| CN103278648B (en) * | 2013-05-24 | 2015-01-21 | 宁波美康盛德医学检验所有限公司 | Albumin detection reagent |
| 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 |
| CN104198686A (en) * | 2014-08-14 | 2014-12-10 | 苏州康铭诚业医用科技有限公司 | Compound stabilizer for uric acid assay kit |
| CN104198686B (en) * | 2014-08-14 | 2017-01-04 | 苏州康铭诚业医用科技有限公司 | A kind of testing uric acid test kit |
| CN104198473B (en) * | 2014-08-14 | 2017-07-07 | 上海睿康生物科技有限公司 | A kind of uric acid detection kit of stabilization |
| CN106404758A (en) * | 2015-07-28 | 2017-02-15 | 珠海和凡医药股份有限公司 | Test paper for detecting uric acid content range in urine |
| CN109916891A (en) * | 2019-04-12 | 2019-06-21 | 吉林省汇酉生物技术股份有限公司 | A kind of dry chemistry reagent piece and preparation method thereof quantitative determining uric acid concentration |
| CN114200122A (en) * | 2021-11-09 | 2022-03-18 | 深圳市锦瑞生物科技股份有限公司 | Freeze-dried reagent ball for uric acid detection, configuration method thereof and microfluidic detection chip |
| CN114200122B (en) * | 2021-11-09 | 2023-09-15 | 深圳市锦瑞生物科技股份有限公司 | Uric acid detection freeze-dried reagent ball, configuration method thereof and microfluidic detection chip |
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Application publication date: 20120418 |