CN105241873B - A kind of fat enzyme detection kit - Google Patents
A kind of fat enzyme detection kit Download PDFInfo
- Publication number
- CN105241873B CN105241873B CN201510582658.2A CN201510582658A CN105241873B CN 105241873 B CN105241873 B CN 105241873B CN 201510582658 A CN201510582658 A CN 201510582658A CN 105241873 B CN105241873 B CN 105241873B
- Authority
- CN
- China
- Prior art keywords
- reagent
- kit
- detection kit
- enzyme detection
- months
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of fatty enzyme detection kits, belong to clinical vitro detection reagent technique field.Kit of the present invention includes reagent R1, reagent R2, calibration object.By adding in the compound stabilizer being made of 4 FPBA, glucomannans, NaCl, propylene glycol, Triton X-100, BSA in reagent R2, effectively raise the stability of kit, its range of linearity is also preferable, and the accuracy of reagent is high, is conducive to further promote the use of in the market.
Description
Technical field
The present invention relates to clinical vitro detection reagent technique field, more particularly to a kind of fatty enzyme detection kit.
Background technology
Lipase (Lipase, glycerol ester hydrolase, LPS) is under the jurisdiction of carboxylic ester hydrolase class, is a kind of hydrolysis of long chain fat
The enzyme of fatty acid glyceride, can be progressively by triglyceride hydrolysis into glycerine and aliphatic acid, be one of digestive ferment of pancreatic secretion.
In addition to serum, lipase active can be also measured in stomach, mucous membrane of small intestine, lung, leucocyte, adipocyte, milk etc..Normal blood
In, only have a small amount of lipase, lipase is easily removed by kidney in blood, and when pancreatic secretion is hyperfunction, ductus pancreaticus is obstructed or pancreas is damaged
Or during necrosis, lipase adverse current or blood is directly accessed, increases in blood lipase activity.Under normal circumstances, LPS is dense in pancreas
Degree is 20000 times of serum, be liver, duodenal 100 times or so.When acute parotidits occur, LPS secretions increase,
The LPS of high concentration is made to enter in blood and is increased in regularity, is shown as：Blood LPS 4~8h after acute parotidits morbidity start
Increase, reach peak value for 24 hours, recover normal after continuing 8~14d, sensitivity：80%~100%, specificity：84%~96%.Serum
LPS expressions, into notable positive correlation, are that the main monitoring of the acute parotidits state of an illness refers to acute parotidits disease progression degree
Mark.
At present, the common LPS assay methods in laboratory have titration, pH-Stat methods, turbidimetry and fluorimetry.Drop
The method of determining is classical way, but this method poor sensitivity, need to use a large amount of serum, the reaction time is long, in addition olive oil emu quality
Quality also directly affects the accuracy of lipase activity measurement result, uses less；PH-Stat methods are laborious, time-consuming because of its, it is difficult to
It automates and needs special instrument and be not widely adopted；There are about the extinctions of 3%-5% in the result of nephelometry measure sample
There is negative test due to molecule is assembled in degree；Fluorimetry is a kind of convenience, special and sensitive lipase activity determination method,
But it is needed equipment costliness, it is difficult to adapt to routine clinical chemical laboratory application.
Fatty enzyme detection kit is based on enzyme development process, using the o- two bays ancestor glycerine -3- glutaric acids-(6'- of 1,2-
Methyl resorufin) -ester should generate methyl resorufin as substrate and lipase reverse, according to the methyl of product at 570nm wavelength
Resorufin generating rate measures lipase active.This method is that one kind need not pre-process sample, and technology and equipment is of less demanding, and
Precision and the higher analysis method of specificity.Since expensive equipment is not required in this method, it can realize automation, and can survey
Fixed a large amount of samples, therefore clinic is subject to be widely popularized.But due to there are enzyme in common lipase enzyme development process detection reagent,
The stability of the reagent can be made to be affected, be unfavorable for the long-term preservation of reagent, so as to cause the bad of poor accuracy and waste
Consequence.
The content of the invention
For problems of the prior art, the present invention provides a kind of fatty enzyme detection kits.The kit with
Conventional kit is compared, and stability is good, and accuracy is high, and the range of linearity is good, and sensitivity for analysis is high, is conducive to reagent clinically
Popularization and application.
The present invention is achieved by the following measures：
A kind of lipase enzyme detection kit, which is characterized in that it includes reagent R1, reagent R2 and calibration object, pilot scale
Agent R1 is formed：
7.2 Tris buffer solutions 100mmol/L of pH
Tauroursodeoxycholic acid 35mmol/L
Sodium chloride 40mmol/L
Reagent R2 is formed：
4.0 tartaric acid buffer 9.5mmol/L of pH
Colipase 500U/L
1,2 o- two bays ancestor glycerine -3- glutaric acids -（6 '-methyl resorufin）-ester 0.2mmol/L
4- formylphenyl boronic acids 0.01%
Glucomannans 0.5-1 mg/mL
NaCl 5mmol/L
Propylene glycol 20mg/mL
Triton X-100 1%
BSA 0.1-1g/L。
The ratios of the reagent R1 and reagent R2 when in use are R1：R2=2：1.
The kit of the present invention carries out on the automatic biochemistry analyzer for have the function of double reagent, and specifically used method is such as
Shown in Fig. 1：
Calibration object used in the present invention is the compound calibration object of Landau company of Britain production.
3 μ l of physiological saline, sample or calibration object are added in, are added after adding 200 μ l preincubates 5min of R1 reagents afterwards
After the reagent R2 reactions 5min of 100 μ l, absorbance, and calculate Δ A/min twice is read.
Beneficial effects of the present invention：
Lipase enzyme development process detection kit provided by the invention, by being added in reagent R2 by 4- formylphenyls
Boric acid（4-FPBA）, glucomannans, NaCl, propylene glycol, Triton X-100, BSA composition compound stabilizer, each component collaboration
Effect makes stable reagent performance excellent, solves enzyme and preserves this unstable problem for a long time, in test it can increase the steady of enzyme
It fixes time, the activity without influencing enzyme, so as to effectively enhance the stability of kit, without the accuracy to reagent
It is had an impact with sensitivity for analysis, is conducive to the reagent and further promotes in the market.
Description of the drawings
Concrete operations schematic diagram of Fig. 1 reagents of the present invention on there is the automatic biochemistry analyzer of double reagent；
2 accuracy validation laboratory test results of Fig. 2 embodiments and control group testing result correlation；
3 accuracy validation laboratory test results of Fig. 3 embodiments and control group testing result correlation；
4 accuracy validation laboratory test results of Fig. 4 embodiments and control group testing result correlation.
Specific embodiment
In order to be better understood from the present invention, it is further described with reference to specific embodiment.
Embodiment 1
A kind of excellent fatty enzyme reagent kit of accuracy of accreditation is obtained in the market, it includes reagent R1, reagent R2, calibration
Product.
Wherein reagent R1 is formed：
7.2 Tris buffer solutions of pH, 100 mmol/L
40 mmol/L of sodium chloride
35 mmol/L of tauroursodeoxycholic acid
Reagent R2 is formed：
4.0 9.5mmol/L of tartaric acid buffer pH
Colipase 500U/L
1,2 o- two bays ancestor glycerine -3- glutaric acids -（6 '-methyl resorufin）0.2 mmol/L of -ester
The kit of the present embodiment description, when in use, assay method are to use the Toshiba 120 with double reagent
Automatic analyzer, operation are as follows：
3 μ l of physiological saline, sample or calibration object are added in, are added after adding 200 μ l preincubates 5min of R1 reagents afterwards
After the reagent R2 reactions 5min of 100 μ l, absorbance, and calculate Δ A/min twice is read.
Calibration object used in the present embodiment is the compound calibration object of Landau company of Britain production.
Embodiment 2
A kind of fat enzyme detection kit, it includes reagent R1, reagent R2, calibration object.
Wherein reagent R1 is formed：
7.2 Tris buffer solutions of pH, 100 mmol/L
40 mmol/L of sodium chloride
35 mmol/L of tauroursodeoxycholic acid
Reagent R2 is formed：
4.0 9.5mmol/L of tartaric acid buffer pH
Colipase 500U/L
1,2 o- two bays ancestor's glycerine -3- penta 2
Acid-（6 '-methyl resorufin）0.2 mmol/L of -ester
4-FPBA 0.01%
0.5 mg/mL of glucomannans
NaCl 5mmol/L
Propylene glycol 20mg/mL
Triton X-100 1%
BSA 0.1g/L
Specific assay method is the same as embodiment 1.
Embodiment 3
A kind of fat enzyme detection kit, it includes reagent R1, reagent R2, calibration object.
Wherein reagent R1 is formed：
7.2 Tris buffer solutions of pH, 100 mmol/L
40 mmol/L of sodium chloride
35 mmol/L of tauroursodeoxycholic acid
Reagent R2 is formed：
4.0 9.5mmol/L of tartaric acid buffer pH
Colipase 500U/L
1,2 o- two bays ancestor's glycerine -3- penta 2
Acid-（6 '-methyl resorufin）0.2 mmol/L of -ester
4-FPBA 0.01%
0.8 mg/mL of glucomannans
NaCl 5mmol/L
Propylene glycol 20mg/mL
Triton X-100 1%
BSA 0.5g/L
Specific assay method is the same as embodiment 1.
Embodiment 4
A kind of fat enzyme detection kit, it includes reagent R1, reagent R2, calibration object.
Wherein reagent R1 is formed：
7.2 Tris buffer solutions of pH, 100 mmol/L
40 mmol/L of sodium chloride
35 mmol/L of tauroursodeoxycholic acid
Reagent R2 is formed：
4.0 9.5mmol/L of tartaric acid buffer pH
Colipase 500U/L
1,2 o- two bays ancestor's glycerine -3- penta 2
Acid-（6 '-methyl resorufin）0.2 mmol/L of -ester
4-FPBA 0.01%
1 mg/mL of glucomannans
NaCl 5mmol/L
Propylene glycol 20mg/mL
Triton X-100 1%
BSA 1g/L
Specific assay method is the same as embodiment 1.
Experimental verification is carried out to kit assay performance obtained in above-described embodiment 1-4.
Accuracy validation is tested：
Using the kit of embodiment 2,3,4 as experimental group, the accuracy for obtaining accreditation in embodiment 1 in the market is excellent
Fatty enzyme reagent kit carries out contrast experiment as a control group, and 40 samples are detected, result such as Fig. 2-Fig. 4 of detection
。
By the detection data of Fig. 2-Fig. 4,2,3,4 detection kit of embodiment and control test kit
Testing result linearly dependent coefficient r is respectively 0.9971,0.9985,0.9981, and correlation is relatively good, shows the reagent of the present invention
Box has high consistency with the fatty enzyme detection kit with excellent accuracy for obtaining accreditation in the market, it was demonstrated that present invention examination
Other various compositions of agent box addition will not impact its accuracy, and kit still keeps preferable accuracy.
Linear dependence confirmatory experiment：
Lipase high level sample is found as 700U/L, is serially diluted with physiological saline, prepares the sample of 6 various concentrations
This, is followed successively by the sample of 700U/L, 560U/L, 420U/L, 280U/L, 140U/L, 0U/L concentration, each each sample of concentration level
It measures respectively three times, takes its average value respectively.The reagent for being utilized respectively embodiment 1,2,3,4 is detected.Testing result such as table 1
It is shown.
1 embodiment 1-4 linear correlation confirmatory experiment testing results of table
| Theoretical concentration（U/L） | 1 testing result of embodiment（U/L） | 2 testing result of embodiment（U/L） | 3 testing result of embodiment（U/L） | 4 testing result of embodiment（U/L） |
| 0 | 3 | 2 | 4 | 4 |
| 140 | 140 | 140 | 139 | 146 |
| 280 | 287 | 274 | 280 | 283 |
| 420 | 425 | 438 | 407 | 398 |
| 560 | 531 | 578 | 571 | 536 |
| 700 | 722 | 698 | 687 | 693 |
| Correlation coefficient r | 0.9980 | 0.9993 | 0.9994 | 0.9992 |
Above-mentioned testing result shows that embodiment 1-4 testing result correlations are all higher than 0.990, but the inspection of embodiment 2,3,4
It surveys result and is more than 0.999, compared with Example 1 with better linear dependence, it is better that this illustrates that reagent of the present invention has
Linear dependence.
Stability confirmatory experiment：
The store reagents in 2 DEG C~8 DEG C, the light protected environment of non-corrosive gas detect the stabilization of four kinds of embodiment reagents
Property.Four kinds of reagents monthly choose same sample measures its absorbances three times, are averaged, and are detected with fresh 1 reagent of embodiment
As a result compared, so that it is determined that the stabilization time of reagent.Detect data such as table 2.
2 stability confirmatory experiment testing result of table
| Time | Embodiment 1 is tried Agent testing result | 2 reagent of embodiment Testing result | 3 reagent of embodiment Testing result | 4 reagent of embodiment Testing result | Fresh embodiment 1 is tried Agent testing result |
| 12 months | 0.01657 | 0.01606 | 0.01687 | 0.01698 | 0.01679 |
| 13 months | 0.01568 | 0.01587 | 0.01646 | 0.01632 | 0.01682 |
| 14 months | 0.01656 | 0.01668 | 0.01654 | 0.01674 | 0.01664 |
| 15 months | 0.01326 | 0.01399 | 0.01387 | 0.01355 | 0.01406 |
| 16 months | 0.00975 | 0.01966 | 0.01967 | 0.01008 | 0.01987 |
| 17 months | 0.00221 | 0.01734 | 0.01686 | 0.01691 | 0.01729 |
| 18 months | 0.00063 | 0.01826 | 0.01757 | 0.01811 | 0.01843 |
| 19 months | 0.00002 | 0.01453 | 0.01432 | 0.01463 | 0.01457 |
| 20 months | 0.00001 | 0.01685 | 0.01596 | 0.01623 | 0.01635 |
| 21 months | 0.00000 | 0.01374 | 0.01421 | 0.01417 | 0.01461 |
| 22 months | 0.00000 | 0.01284 | 0.01314 | 0.01321 | 0.01358 |
| 23 months | 0.00000 | 0.01263 | 0.01198 | 0.01167 | 0.01265 |
| 24 months | 0.00000 | 0.01198 | 0.01213 | 0.01191 | 0.01235 |
| 25 months | 0.00000 | 0.00857 | 0.00843 | 0.00830 | 0.01232 |
| 26 months | 0.00000 | 0.00065 | 0.00082 | 0.00097 | 0.01328 |
Experimental result shows, 1 reagent of embodiment is stored 15 months in 2 DEG C~8 DEG C, the light protected environment of non-corrosive gas
Stablize, and 2,3,4 reagent of embodiment stores 24 months stabilizations in 2 DEG C~8 DEG C, the light protected environment of non-corrosive gas, explanation
In reagent the stability of fatty enzyme detection kit can be effectively improved by adding in compound stabilizer.
In summary analyze, fat enzyme detection kit provided by the invention, by adding in stable composition in reagent R2
Agent can effectively improve the stability of kit, and the range of linearity is preferable, and the accuracy of reagent is also preferable.Therefore, the present invention carries
The fatty enzyme detection kit supplied is conducive to further promote the use of in the market.
Claims (2)
1. a kind of fat enzyme detection kit, which is characterized in that it includes reagent R1, reagent R2 and calibration object, wherein reagent R1
It forms and is：
7.2 Tris buffer solutions 100mmol/L of pH
Tauroursodeoxycholic acid 35mmol/L
Sodium chloride 40mmol/L
Reagent R2 is formed：
4.0 tartaric acid buffer 9.5mmol/L of pH
Colipase 500U/L
1,2 o- two bays ancestor glycerine -3- glutaric acids -（6 '-methyl resorufin）-ester 0.2mmol/L
4- formylphenyl boronic acids 0.01%
Glucomannans 0.5-1 mg/mL
NaCl 5mmol/L
Propylene glycol 20mg/mL
Triton X-100 1%
BSA 0.1-1g/L。
2. kit according to claim 1, which is characterized in that the reagent R1 and the ratios of reagent R2 when in use are
R1：R2=2：1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510582658.2A CN105241873B (en) | 2015-09-14 | 2015-09-14 | A kind of fat enzyme detection kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510582658.2A CN105241873B (en) | 2015-09-14 | 2015-09-14 | A kind of fat enzyme detection kit |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105241873A CN105241873A (en) | 2016-01-13 |
| CN105241873B true CN105241873B (en) | 2018-05-22 |
Family
ID=55039596
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510582658.2A Active CN105241873B (en) | 2015-09-14 | 2015-09-14 | A kind of fat enzyme detection kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105241873B (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107782680A (en) * | 2016-08-26 | 2018-03-09 | 山东博科生物产业有限公司 | A kind of antiheparin fat enzyme detection kit of stabilization |
| CN107782683A (en) * | 2016-08-27 | 2018-03-09 | 山东博科生物产业有限公司 | A kind of glutamate dehydrogenase enzyme detection kit |
| CN108169152A (en) * | 2017-12-27 | 2018-06-15 | 山东博科生物产业有限公司 | A kind of angiotensin converting enzyme detection kit and its application method |
| CN109837270B (en) * | 2018-06-19 | 2022-09-09 | 深圳市安帝宝科技有限公司 | A method for stabilizing inositol dehydrogenase, ketoamine oxidase and sphingomyelinase in liquid for long time |
| CN109212176B (en) * | 2018-08-30 | 2019-10-11 | 中拓生物有限公司 | A kind of pyruvic acid assay kit and its preparation method and application |
| CN109298190B (en) * | 2018-09-26 | 2020-04-28 | 山东博科生物产业有限公司 | High density lipoprotein cholesterol detect reagent box |
| CN110923292B (en) * | 2019-11-15 | 2024-03-29 | 中山市创艺生化工程有限公司 | Serum lipase detection kit and preparation method and application thereof |
| CN113008812B (en) * | 2021-01-05 | 2022-04-08 | 中元汇吉生物技术股份有限公司 | Kit for quantitatively detecting lipase LPS |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1909796A (en) * | 2004-01-30 | 2007-02-07 | 巴斯福股份公司 | Stabilized phosphatase formulations |
| CN104198474A (en) * | 2014-08-14 | 2014-12-10 | 上海睿康生物科技有限公司 | Detection kit for measuring content of lipase in serum by colorimetric method |
| CN104215632A (en) * | 2014-08-28 | 2014-12-17 | 宁波瑞源生物科技有限公司 | Stable lipase kit |
| CN104745673A (en) * | 2015-04-28 | 2015-07-01 | 山东博科生物产业有限公司 | Stable 5 minute - ribonucleotide hydrolytic enzyme detection kit |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7514255B2 (en) * | 2004-12-03 | 2009-04-07 | Idexx Laboratories, Inc. | Methods and devices for detecting pancreatic lipase |
-
2015
- 2015-09-14 CN CN201510582658.2A patent/CN105241873B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1909796A (en) * | 2004-01-30 | 2007-02-07 | 巴斯福股份公司 | Stabilized phosphatase formulations |
| CN104198474A (en) * | 2014-08-14 | 2014-12-10 | 上海睿康生物科技有限公司 | Detection kit for measuring content of lipase in serum by colorimetric method |
| CN104215632A (en) * | 2014-08-28 | 2014-12-17 | 宁波瑞源生物科技有限公司 | Stable lipase kit |
| CN104745673A (en) * | 2015-04-28 | 2015-07-01 | 山东博科生物产业有限公司 | Stable 5 minute - ribonucleotide hydrolytic enzyme detection kit |
Non-Patent Citations (1)
| Title |
|---|
| GPIHBP1 stabilizes lipoprotein lipase and prevents its inhibition by angiopoietin-like 3 and angiopoietin-like 4;William K. Sonnenburg 等;《Journal of Lipid Research》;20090621;第50卷;2421-2429 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105241873A (en) | 2016-01-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105241873B (en) | A kind of fat enzyme detection kit | |
| CN102721684B (en) | Two-step enzyme measuring method and measuring reagent for creatinine in blood serum | |
| US20080038765A1 (en) | Composition for Lipase Activity Determination and Method of Determing Activity | |
| CN109239059A (en) | A kind of glycated serum protein assay kit and its preparation method and application | |
| CN104198473A (en) | Stable uric acid detection kit | |
| CN104198474A (en) | Detection kit for measuring content of lipase in serum by colorimetric method | |
| CN101226198A (en) | Enzymatical detection method of saccharify blood albumen as well as liquid stabilising agent | |
| CN111808921A (en) | Trinder reaction-based detection kit and application thereof | |
| CN110261625A (en) | A kind of bilirubin multiplexed detection reagents box and its application method | |
| CN112051354B (en) | Lipase determination kit and preparation method thereof | |
| CN104714036A (en) | Preparation method of antithrombase III activity determination kit | |
| CN104198407A (en) | Detection kit for detecting content of beta-hydroxybutyrate in serum by adopting stable enzymatic method | |
| CN104048929A (en) | Detection kit for total bilirubin | |
| CN114200122B (en) | Uric acid detection freeze-dried reagent ball, configuration method thereof and microfluidic detection chip | |
| CN103266165B (en) | Amylase detection reagent | |
| CN112575057B (en) | Composition and application thereof in detecting activity of lipoprotein-associated phospholipase A2 | |
| CN104048928A (en) | Detection kit for direct bilirubin | |
| CN110923292B (en) | Serum lipase detection kit and preparation method and application thereof | |
| CN107356544B (en) | Lipase detect reagent box | |
| CN109490227A (en) | A kind of highly sensitive fatty enzyme reagent kit | |
| CN107703288A (en) | Improve the bile acid detection reagent of reaction stability | |
| CN107782680A (en) | A kind of antiheparin fat enzyme detection kit of stabilization | |
| CN104048955A (en) | High density lipoprotein cholesterol detection kit | |
| CN106324234B (en) | Neu 5 Ac aldolase of modification and its preparation method and application | |
| CN109298190A (en) | A kind of high-density lipoprotein cholesterol detection kit |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |