CN109580504A - A kind of lipoprotein cholesterol measurement reagent and kit - Google Patents
A kind of lipoprotein cholesterol measurement reagent and kit Download PDFInfo
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Abstract
The present invention provides a kind of lipoprotein cholesterol measurement reagent and kit, belong to biochemistry detection technical field, it further include cholesterol esterase, peroxidase, catalase, hydrogen peroxide enzyme inhibitor, surfactant, emulsifier, polyanion, bivalent cation, stabilizer, anti-interference agent, color developing agent and preservative containing poly- а-alkene, cholesterol oxidase and reaction promoter.The present invention can be used for automatic clinical chemistry analyzer detection, easy to operate, and detection efficiency and accuracy can be improved, reduce testing cost.
Description
Technical field
The invention belongs to biochemistry detection technical fields, and in particular to a kind of lipoprotein cholesterol measurement reagent and kit.
Background technique
According to the difference of physical property, human plasma lipoprotein can be divided mainly into following four: high-density lipoprotein
(HDL), low-density lipoprotein (LDL), very low density lipoprotein (VLDL) and chylomicron (CM), they are cholesterol in blood plasma
Be primarily present and types of transportation.Wherein high-density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C)
Content respectively to the incidence of human atherosclerotic class disease coronary heart disease at negative, positive related, the two containing in human plasma
Affected by many factors, such as age, gender, inherent cause, smoking/motion conditions, diet, obesity are measured, by China's " blood
Rouge exception remedial proposal " propose judgment criteria, by China " dyslipidemia remedial proposal " proposition judgment criteria, normal condition
Under, the HDL-C of male is between 1.16-1.42mmol/L, and women is between 1.29-1.55mmol/L；The average LDL-C of young people contains
Amount is about 2.7mmol/L, and the middle-aged and the old is about 3.37mmol/L.The measurement of HDL-C, LDL-C content is of great significance, and is mesh
The important risk factor index of preceding clinically cardiovascular and cerebrovascular disease.
There are many measuring methods of HDL-C, LDL-C content, after ultracentrifugation, electrophoresis, measurement side in the market
Method is all direct measuring method.The direct measuring method of HDL-C, LDL-C can all be divided into three generations, be made extensively by each clinical labororatory at present
It is the even phase measuring method of the third generation, this method is all two-step method, it is not required to sample preprocessing, precipitation and separation etc. and artificially handles,
Reagent and sample dosage are few, can measure on automatic clinical chemistry analyzer, and the accuracy of measurement result and accuracy are higher.HDL-
The even phase measuring method of C mainly include following 5 kinds: PEG/ antibody pack (IRC), PEG enzyme modification method (PEGME), reaction promoter/
Peroxidase removes method (SPD), catalase removes method (CAT), antibody mediated immunity partition method (AB).The even phase measurement of LDL-C
Method is main are as follows: soluble reaction method (SOL), surfactant remove method (SUR), protectiveness reagent method (PRO), catalase
Removing method (CAT), calixarenes method (CAL).
Different measuring method principles are different, but all generally existing certain defect, in first step reaction cannot completely by
Non-HDL-C or non-LDL-C package is removed, and it is higher to easily lead to measurement result；Second step reaction in HDL-C or LDL-C not
It can discharge completely, it is relatively low to easily lead to measurement result；In blood plasma phosphatide, cholesterol exception patient's sample in, HDL-C or LDL-
C measurement will appear exception, and anti-interference ability is weak, and stability is poor.
Summary of the invention
The object of the present invention is to provide a kind of lipoprotein cholesterol measurement reagent and kits, can be used for full-automatic biochemical point
Analyzer detection, it is easy to operate, detection efficiency and accuracy can be improved, reduce testing cost.
The present invention provides the following technical solutions:
A kind of lipoprotein cholesterol measurement reagent, contains poly- а-alkene, cholesterol oxidase and reaction promoter.
Preferably, the concentration of the а-alkene is 0.0005-0.005% (v/v).
Preferably, the molecular weight of the cholesterol oxidase is 25-45KDa, concentration 0.1-2KU/L.
Preferably, the reaction promoter is Flufenamic acid, mefenamic acid, Fusidic Acid, bromo- 2,2:6, the 2- tri- of 6,6- bis-
At least one of bipyridyl, coban, concentration 50-200umol/L.
It preferably, further include cholesterol esterase, peroxidase, catalase, hydrogen peroxide enzyme inhibitor, surface work
Property agent, emulsifier, polyanion, bivalent cation, stabilizer, anti-interference agent, color developing agent and preservative.
Preferably, cholesterol esterase is at least one of modification enzyme or unmodified enzyme, concentration 0.1-2KU/L.
Preferably, the modifier of enzyme is at least one of PEG6000, both arms PEG7000, diethylenetriamine pentaacetic acid.
Preferably, the concentration of peroxidase is 0.1-3KU/L.
Preferably, the concentration of catalase is 0.1-3KU/L.
Preferably, hydrogen peroxide enzyme inhibitor is Sodium azide, 3- amino -1,2, one or two kinds of, concentration in 4- triazole
For 1-30g/L.
Preferably, the surfactant is Emulgen B-66, Triton-X, trimethyl-β-cyclodextrin, ethylene oxide
Octadecylamine, poloxamer-F88, Bu Lijie -58, TWEEN Series, SDS, ether carboxylate, alkyl glycerylether, polyoxyethylene cetyl
At least one of base ether, NONIN HS 240, the HLB value of the surfactant are 10-15.
Preferably, emulsifier is at least one of ethyl alcohol, fatty alcohol polyoxyethylene ether, polyethenoxy ether.
Preferably, polyanion is heparin sulfate, dextran sulfate, tungstophosphoric acid sodium, at least one in polyethylene glycol
Kind, concentration 0.1-1g/L.
Preferably, the bivalent cation is Mg2+、Mn2+、Ca2+At least one of, concentration 0.1-5g/L.
Preferably, the stabilizer can be glutathione, trehalose, lecithin, glycerol, EDTA, glycine betaine, gelatin, ox
At least one of haemocyanin, concentration 0.1-5g/L.
Preferably, anti-interference agent be at least one of chloroform, protamine sulfate, mannitol, ascorbic acid oxidase,
Concentration is 0.1-6g/L.
Preferably, color developing agent is phenol, sodium 3,5-dichloro-2-hydroxybenzenesulfonate (DHBS), N- ethyl-N- (2- hydroxyl -3-
Third sulfo group) meta-aminotoluene (TOOS), N, N- bis- (4- sulphur butyl) -3- methylaniline disodium (TODB), N- ethyl-N- (3- sulphur third
At least one of base) -3- methylaniline sodium salt (TOPS), concentration 5-15mL/L.
Preferably, preservative is Sodium azide, gentamicin, Proclin series, at least one of KroVin series, dense
Degree is 1-15g/L.
A kind of kit for HDL-C measurement, including the first reagent and the second reagent, first reagent contain buffering
Liquid, cholesterol oxidase, peroxidase, surfactant, anti-interference agent, reaction promoter, preservative, bivalent cation,
Stabilizer and polyanion, second reagent contain buffer, catalase, surfactant, anti-interference agent, poly- а-
Alkene, preservative, stabilizer, color developing agent and 4-AA.
A kind of kit for LDL-C measurement, including the first reagent and the second reagent, first reagent contain buffering
Liquid, poly- а-alkene, cholesterol oxidase, cholesterol esterase, catalase, surfactant, anti-interference agent, reaction promote
Agent, preservative and stabilizer, second reagent contain buffer, color developing agent, 4-AA, peroxidase, table
Face activating agent, preservative, hydrogen peroxide enzyme inhibitor and stabilizer.
The beneficial effects of the present invention are:
(1) lipoprotein cholesterol provided by the invention measures reagent, introduces poly- а-alkene to improve lipoprotein cholesterol survey
Fixed accuracy, poly- а-alkene can be improved the selectivity of surfactant.
(2) lipoprotein cholesterol provided by the invention measures reagent, introduces the cholesterol oxidase of low molecular weight, Neng Gouti
The Scavenging activity of high lipoprotein cholesterol further increases the accuracy of different lipoprotein cholesterol measurements, such cholesterol oxygen
Changing enzyme can be improved the oxidation of cholesterol in reaction.
(3) lipoprotein cholesterol provided by the invention measures reagent, introduces one or more kinds of reaction promoters to mention
High lipoprotein cholesterol determination efficiency, on the one hand can reduce the dosage of enzyme, save the cost, on the other hand, to a certain extent
The reaction time can be shortened.
(4) kit of HDL-C measurement provided by the invention, poly- а-alkene of addition improve surfactant to HDL-
The specificity of C improves the accuracy of HDL-C measurement.
(5) kit of LDL-C measurement provided by the invention, poly- а-alkene of addition improve surfactant to LDL-
The selection of C improves the accuracy of LDL-C measurement.
Detailed description of the invention
Attached drawing is used to provide further understanding of the present invention, and constitutes part of specification, with reality of the invention
It applies example to be used to explain the present invention together, not be construed as limiting the invention.In the accompanying drawings:
Fig. 1 is the linear dependence figure of embodiment 2 kit test value and theoretical value in the test of embodiment 21；
Fig. 2 is the linear dependence figure of 2 kit of embodiment and contrast agents box in the test of embodiment 24；
Fig. 3 is the linear dependence figure of embodiment 3 kit test value and theoretical value in the test of embodiment 31；
Fig. 4 is the linear dependence figure of 3 kit of embodiment and contrast agents box in the test of embodiment 34.
Specific embodiment
Embodiment 1
A kind of lipoprotein cholesterol measurement reagent, contains poly- а-alkene, cholesterol oxidase and reaction promoter.The а-
The concentration of alkene is 0.0005-0.005% (v/v).The molecular weight of the cholesterol oxidase is 25-45KDa, concentration 0.1-
2KU/L.The reaction promoter is Flufenamic acid, mefenamic acid, Fusidic Acid, the bromo- 2,2:6,2- terpyridyl of 6,6- bis-, not
At least one of energy rhzomorph, concentration 50-200umol/L.
It further includes cholesterol esterase, peroxidase, catalase, hydrogen peroxide that the lipoprotein cholesterol, which measures reagent,
Enzyme inhibitor, surfactant, emulsifier, polyanion, bivalent cation, stabilizer, anti-interference agent, color developing agent and anti-corrosion
Agent.
Cholesterol esterase (CHE) is at least one of modification enzyme or unmodified enzyme, concentration 0.1-2KU/L.Enzyme is repaired
Jewelry is at least one of PEG6000, both arms PEG7000, diethylenetriamine pentaacetic acid.
Peroxidase (POD) may originate from plant (such as soybean, radish), animal (such as animal leucocyte), microorganism (such as ferment
At least one of it is female), concentration 0.1-3KU/L.
Catalase (CAT) may originate from plant (chloroplaset), animal (such as cattle liver), microorganism (such as red pseudomonas
At least one of belong to), concentration 0.1-3KU/L.
Hydrogen peroxide enzyme inhibitor is Sodium azide, 3- amino -1,2, one or two kinds of, concentration 1-30g/ in 4- triazole
L。
Surfactant is Emulgen B-66, Triton-X, trimethyl-β-cyclodextrin, ethylene oxide octadecylamine, moors
Luo Shamu-F88, Bu Lijie -58, TWEEN Series, SDS, ether carboxylate, alkyl glycerylether, polyoxyethylene cetyl base ether, polyoxy second
At least one of alkene octyl phenyl ether, the HLB value of the surfactant are 10-15.
Emulsifier is at least one of ethyl alcohol, fatty alcohol polyoxyethylene ether, polyethenoxy ether.
Polyanion is at least one of heparin sulfate, dextran sulfate, tungstophosphoric acid sodium, polyethylene glycol, concentration
For 0.1-1g/L.
Bivalent cation is Mg2+、Mn2+、Ca2+At least one of, concentration 0.1-5g/L.
Stabilizer can be in glutathione, trehalose, lecithin, glycerol, EDTA, glycine betaine, gelatin, bovine serum albumin
At least one, concentration 0.1-5g/L.
Anti-interference agent is at least one of chloroform, protamine sulfate, mannitol, ascorbic acid oxidase, and concentration is
0.1-6g/L。
Color developing agent is phenol, the chloro- 2- sodium hydroxybenzenesulfonate (DHBS) of 3,5- bis-, N- ethyl-N- (the third sulfo group of 2- hydroxyl -3-)
Meta-aminotoluene (TOOS), N, N- bis- (4- sulphur butyl) -3- methylaniline disodium (TODB), N- ethyl-N- (3- sulfopropyl) -3- first
At least one of base aniline sodium salt (TOPS), concentration 5-15mL/L.
Preservative is at least one of Sodium azide, gentamicin, Proclin series, KroVin series, concentration 1-
15g/L。
Embodiment 2
A kind of kit for HDL-C measurement, including the first reagent and the second reagent, first reagent contain buffering
Liquid, cholesterol oxidase, peroxidase, surfactant, anti-interference agent, reaction promoter, preservative, bivalent cation,
Stabilizer and polyanion, second reagent contain buffer, catalase, surfactant, anti-interference agent, poly- а-
Alkene, preservative, stabilizer, color developing agent and 4-AA.
The specific raw material and proportion of this kit are as follows:
The composition and concentration of 1 first reagent of table
| Buffer PIPES (PH=7.0) | 50mmol/L |
| Cholesterol oxidase (25-45KDa, CHO) | 1.5KU/L |
| Cholesterol esterase (CHE) | 1.5KU/L |
| Peroxidase (POD) | 1.5KU/L |
| Bu Lijie -58 | 0.5% |
| Ethylene oxide octadecylamine | 0.5% |
| Protamine sulfate | 3g/L |
| Ascorbic acid oxidase (ASO) | 2KU/L |
| Flufenamic acid | 120umol/L |
| Proclin 300 | 5g/L |
| MgSO4 | 2g/L |
| Glycerol | 8ml/L |
| Dextran sulfate (DS) | 0.5g/L |
| Alpha-cyclodextrin | 0.5g/L |
The composition and concentration of 2 second reagent of table
| Buffer PIPES (PH=7.0) | 50mmol/L |
| Catalase (CAT) | 1.5KU/L |
| Emulgen B-66 | 1% |
| Mannitol | 5g/L |
| Poly- а-alkene | 0.001% |
| Proclin 300 | 5g/L |
| Glycine betaine | 0.3mol/L |
| TOOS | 8mL/L |
| 4-AA (4-AAP) | 1g/L |
The present embodiment selects the SPD method in even phase measuring method, and this method is based on following principle: using surfactant and not
With lipoprotein affinity difference, directly measurement HDL-C.This method mainly includes two-step reaction: in first step reaction, utilizing reaction
Promotor (polymer/surfactant of synthesis) makes non-HDL form soluble complex, and under POD effect, makes multiple
The free cholesterol in object surface is closed to be removed；In second step reaction, under Action of Surfactant, keeps HDL particle solvable, release
The cholesterol put generates livid purple color benzoquinones pigment with amino-antipyrine under CHO, CHE and CAT effect.By measuring absorbance
Realize the purpose that HDL-C is directly measured without precipitation and separation.
The present embodiment detects HDL-C in sample, and sample to be tested is one of serum, blood plasma, and specific steps are such as
Under:
(1) the first reagent of 300uL and 3uL sample to be tested are uniformly mixed, obtain reaction solution 1 after 37 DEG C of incubations；
(2) the second reagent of 100uL is then added to be uniformly mixed, obtains reaction solution 2, measures the absorbance A 1 of reaction solution 2；
(3) absorbance A 2 of reaction solution 2 is measured after 5min again, measurement dominant wavelength is 600nm, a length of 700nm of complementary wave；
(4) absorbance of high-density lipoprotein cholesterol titer is measured with step (1)-(3) operation, respectively
Absorbance A 3 and A4 are obtained, measurement dominant wavelength is 600nm, a length of 700nm of complementary wave；
(5) it is calculated with absorbance difference, HDL-C content: HDL-C concentration=sample absorbance is calculated according to formula
(A2-A1) × concentration of standard solution/titer absorbance (A4-A3).
Experimental example
Using other company's similar-type products as contrast agents box, with the examination provided in this embodiment for HDL-C measurement
Agent box carries out the test of HDL-C simultaneously, and the results are shown in Table 3:
3 embodiment of table, 2 kit and contrast agents box parameter comparison
| The range of linearity (mmol/L) | Accuracy | Sensitivity for analysis (mmol/L) | |
| Contrast agents box | 0.00~3.00 | ≤ ± 10% | 3.0 |
| 2 kit of embodiment | 0.00~5.50 | ≤ ± 10% | 1.5 |
Contrast agents box is a domestic commercially available HDL-C detection kit, ingredient and formula are as follows: reagent 1 includes PIPES
Buffer, TOOS, ascorbic acid oxidase, Tween 80；Reagent 2 includes PIPES buffer, 4-AAP, CHOD, CHER, POD.By
Table 3 is it is found that the sensitivity for analysis of 2 kit of embodiment is higher.
Test 1
Prepare high-density lipoprotein cholesterol various concentration master sample, respectively 0,0.5,1.0,1.5,2.0,2.5,
3.0,3.5,4.0,5,5.5mmol/L measure its concentration with 2 kit of embodiment and contrast agents box respectively, it is all test into
3 repetitions of row, are averaged.As a result such as table 4:
The comparison of 4 range of linearity of table
By table 4 and Fig. 1 result it can be seen that when concentration of specimens Wei≤3.5mmol/L, since the range of linearity is smaller, control
Kit becomes larger surveying timing error, cannot accurate detection HDL-C concentration, and 2 kit of embodiment is still in normal
Error range within, it can be seen that the kit range of linearity of the present invention is wider.
Test 2
Using contrast agents box and 2 kit of embodiment as material, calibrated using Landau calibration object, with Roche high level and
Low value Quality Control is measured, and all tests carry out 3 repetitions, is averaged.As a result such as table 5:
5 accuracy of table comparison
| Target value | Contrast agents box | 2 kit of embodiment |
| High level 1.53mmol/L | 1.56 | 1.52 |
| Low value 0.844mmol/L | 0.85 | 0.84 |
Table 5 the result shows that: for the test result of 2 kit of embodiment closer to target value, accuracy is higher than control kit.
Test 3
Withinrun precision: using Roche high level and low value quality-control product as sample, replication 10 times (n=10), is calculated respectively
The coefficient of variation (CV%) value.Specific such as table 6:
The comparison of 6 withinrun precision of table
By the result of upper table it can be seen that the withinrun precision of 2 kit of embodiment is higher than contrast agents box.
Test 4
Calibrated with Landau calibration object, measure the content of the HDL-C of 40 clinical samples, such as table 7 and Fig. 2 the result shows that
2 kit test result of embodiment has good correlation with contrast agents acquired results, and all tests carry out 3 repetitions.
The comparison of 7 linear dependence of table
Embodiment 3
A kind of kit for LDL-C measurement, including the first reagent and the second reagent, first reagent contain buffering
Liquid, poly- а-alkene, cholesterol oxidase, cholesterol esterase, catalase, surfactant, anti-interference agent, reaction promote
Agent, preservative and stabilizer, second reagent contain buffer, color developing agent, 4-AA, peroxidase, table
Face activating agent, preservative, hydrogen peroxide enzyme inhibitor and stabilizer.
The specific raw material and proportion of this kit are as follows:
The composition and concentration of 8 first reagent of table
The composition and concentration of 9 second reagent of table
| Buffer MOPS (PH=7.0) | 50mmol/L |
| TOOS color developing agent | 10mL/L |
| 4-AA (4-AAP) | 1g/L |
| Peroxidase (POD) | 1KU/L |
| Triton-X | 1.2% |
| Proclin 300 | 5g/L |
| Sodium azide | 2g/L |
| Glycine betaine | 1.5g/L |
The present embodiment selects the CAT method in even phase measuring method, and this method is based on following principle: using surfactant and not
With lipoprotein affinity difference, directly measurement LDL-C.This method mainly includes two-step reaction: in first step reaction, utilizing surface
Activating agent and ion buffer specific effect discharge the cholesterol of non-LDL in non-LDL, and under CAT effect, make
The cholesterol of release is removed；In second step reaction, under another Action of Surfactant, release the cholesterol of LDL particle
It puts, the cholesterol of release generates livid purple color benzoquinones pigment with amino-antipyrine under CHO, CHE and POD effect.Pass through survey
Determine absorbance and realizes the purpose for directly measuring LDL-C without precipitation and separation.
The present embodiment detects LDL-C in sample, and sample to be tested is one of serum, blood plasma, and specific steps are such as
Under:
(1) the first reagent of 300uL reagent and 3uL sample to be tested are uniformly mixed, obtain reaction solution 1 after 37 DEG C of incubations；
(2) the second reagent of 100uL reagent is then added to be uniformly mixed, obtains reaction solution 2, measures the absorbance of reaction solution 2
A1；
(3) absorbance A 2 of reaction solution 2 is measured after 5min again, measurement dominant wavelength is 545nm, a length of 700nm of complementary wave；
(4) absorbance of high-density lipoprotein cholesterol titer is measured with step (1)-(3) operation, respectively
Absorbance A 3 and A4 are obtained, measurement dominant wavelength is 545nm, a length of 700nm of complementary wave；
(5) it is calculated with absorbance difference, LDL-C content: LDL-C concentration=sample absorbance is calculated according to formula
(A2-A1) × concentration of standard solution/titer absorbance (A4-A3).
Experimental example
Using other company's similar-type products as contrast agents box, with the examination provided in this embodiment for LDL-C measurement
Agent box carries out the test of LDL-C simultaneously, and the results are shown in Table 10:
10 embodiment of table, 3 kit and contrast agents box parameter comparison
| The range of linearity (mmol/L) | Accuracy | Sensitivity for analysis (mmol/L) | |
| Contrast agents box | 0.00~6.00 | ≤ ± 10% | 4.0 |
| 3 kit of embodiment | 0.00~16.00 | ≤ ± 10% | 2 |
Contrast agents box is a domestic commercially available LDL-C detection kit, ingredient and formula are as follows: reagent 1 includes Good '
S buffer；Reagent 2 includes Good ' s buffer, 4-AAP, CHOD, CHER, POD.As shown in Table 10, embodiment 3 kit
Sensitivity for analysis is higher
Test 1
Prepare low density lipoprotein cholesterol various concentration master sample, respectively 0,2,4,6,8,10,12,14,
16mmol/L, measures its concentration with 3 kit of embodiment and contrast agents box respectively, and all tests carry out 3 repetitions, are averaged
Value.As a result such as table 11:
The comparison of 11 range of linearity of table
By table 11 and Fig. 3 result it can be seen that when concentration of specimens Wei≤6mmol/L, since the range of linearity is smaller, control examination
Agent box becomes larger surveying timing error, cannot accurate detection LDL-C concentration, and 3 kit of embodiment is still in normal
Within error range, it can be seen that the kit range of linearity of the present invention is wider.
Test 2
Using contrast agents box and 3 kit of embodiment as material, calibrated using Landau calibration object, with Roche high level and
Low value Quality Control is measured, and all tests carry out 3 repetitions, is averaged.As a result such as table 12:
12 accuracy of table comparison
| Target value | Contrast agents box | 3 kit of embodiment |
| High level 2.62mmol/L | 2.55 | 2.64 |
| Low value 1.56mmol/L | 1.43 | 1.53 |
The result shows that: for the test result of 3 kit of embodiment closer to target value, accuracy is higher than control kit.
Test 3
Withinrun precision: using Roche high level and low value quality-control product as sample, replication 10 times (n=10), is calculated respectively
The coefficient of variation (CV%) value.Specific such as table 13:
The comparison of 13 withinrun precision of table
By the result of upper table it can be seen that the withinrun precision of 3 kit of embodiment is higher than contrast agents box.
Test 4
It is calibrated with Landau calibration object, measures the content of the LDL-C of 40 clinical samples, such as the following table 14 and Fig. 4 result
Show that 3 kit test result of embodiment and contrast agents acquired results have good correlation, all tests carry out 3 weights
It is multiple, it is averaged.
The comparison of 14 linear dependence of table
The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention, although referring to aforementioned reality
Applying example, invention is explained in detail, for those skilled in the art, still can be to aforementioned each implementation
Technical solution documented by example is modified or equivalent replacement of some of the technical features.It is all in essence of the invention
Within mind and principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of lipoprotein cholesterol measures reagent, which is characterized in that promote containing poly- а-alkene, cholesterol oxidase and reaction
Agent.
2. a kind of lipoprotein cholesterol according to claim 1 measures reagent, which is characterized in that the concentration of the а-alkene
For 0.0005-0.005% (v/v).
3. a kind of lipoprotein cholesterol according to claim 1 measures reagent, which is characterized in that the cholesterol oxidase
Molecular weight be 25-45KDa, concentration 0.1-2KU/L.
4. a kind of lipoprotein cholesterol according to claim 1 measures reagent, which is characterized in that the reaction promoter is
At least one of Flufenamic acid, mefenamic acid, Fusidic Acid, bromo- 2,2:6, the 2- terpyridyl of 6,6- bis-, coban are dense
Degree is 50-200umol/L.
5. a kind of lipoprotein cholesterol according to claim 1 measures reagent, which is characterized in that further include cholesteryl ester
Enzyme, peroxidase, catalase, hydrogen peroxide enzyme inhibitor, surfactant, emulsifier, polyanion, divalent sun
Ion, stabilizer, anti-interference agent, color developing agent and preservative.
6. a kind of lipoprotein cholesterol according to claim 5 measures reagent, which is characterized in that the surfactant is
Emulgen B-66, Triton-X, trimethyl-β-cyclodextrin, ethylene oxide octadecylamine, poloxamer-F88, Bu Lijie-
58, tween, SDS, ether carboxylate, alkyl glycerylether, polyoxyethylene cetyl base ether, at least one in NONIN HS 240
Kind, the HLB value of the surfactant is 10-15.
7. a kind of lipoprotein cholesterol according to claim 5 measures reagent, which is characterized in that the bivalent cation is
Mg2+、Mn2+、Ca2+At least one of, concentration 0.1-5g/L.
8. a kind of lipoprotein cholesterol according to claim 5 measures reagent, which is characterized in that the stabilizer can be paddy
At least one of the sweet peptide of Guang, trehalose, lecithin, glycerol, EDTA, glycine betaine, gelatin, bovine serum albumin, concentration 0.1-
5g/L。
9. a kind of kit for HDL-C measurement, which is characterized in that including the first reagent and the second reagent, first examination
Agent contains buffer, cholesterol oxidase, peroxidase, surfactant, anti-interference agent, reaction promoter, preservative, two
Valence cation, stabilizer and polyanion, second reagent contain buffer, catalase, surfactant, resist and do
Disturb agent, poly- а-alkene, preservative, stabilizer, color developing agent and 4-AA.
10. a kind of kit for LDL-C measurement, which is characterized in that including the first reagent and the second reagent, first examination
Agent contain buffer, poly- а-alkene, cholesterol oxidase, cholesterol esterase, catalase, surfactant, anti-interference agent,
Reaction promoter, preservative and stabilizer, second reagent contain buffer, color developing agent, 4-AA, peroxidating
Object enzyme, surfactant, preservative, hydrogen peroxide enzyme inhibitor and stabilizer.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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| CN115932239A (en) * | 2022-12-22 | 2023-04-07 | 广州市进德生物科技有限公司 | Oxidized low-density lipoprotein protective agent, detection kit for determining oxidized low-density lipoprotein and application of detection kit |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110791549A (en) * | 2019-12-03 | 2020-02-14 | 宁波普瑞柏生物技术股份有限公司 | Method and kit for quantitative determination of small dense low density lipoprotein cholesterol |
| CN110791549B (en) * | 2019-12-03 | 2023-09-26 | 宁波普瑞柏生物技术股份有限公司 | Method and kit for quantitatively determining small dense low density lipoprotein cholesterol |
| CN111190017A (en) * | 2019-12-11 | 2020-05-22 | 深圳爱信生物技术有限公司 | Sensitization blocking buffer solution and application thereof |
| CN112014338A (en) * | 2020-08-28 | 2020-12-01 | 武汉生之源生物科技股份有限公司 | Paraoxonase-1 activity detection kit and application thereof |
| CN112695071A (en) * | 2020-12-16 | 2021-04-23 | 浙江伊利康生物技术有限公司 | High-density lipoprotein 3 determination reagent, method and kit |
| CN113740539A (en) * | 2021-08-11 | 2021-12-03 | 重庆中元汇吉生物技术有限公司 | Kit for determining specific growth factor |
| CN114134203A (en) * | 2021-11-26 | 2022-03-04 | 深圳市雷诺华科技实业有限公司 | Method for measuring high-density lipoprotein cholesterol by using nano enzyme |
| CN114216871A (en) * | 2021-12-15 | 2022-03-22 | 青岛汉唐生物科技有限公司 | Compound stabilizer for replacing sodium cholate in cholesterol kit |
| CN114216871B (en) * | 2021-12-15 | 2024-02-02 | 青岛汉唐生物科技有限公司 | Composite stabilizer for substituting sodium cholate in cholesterol kit |
| CN114350744A (en) * | 2022-01-21 | 2022-04-15 | 广西康柏莱科技有限公司 | Small and dense low-density lipoprotein cholesterol detection kit |
| CN115932239A (en) * | 2022-12-22 | 2023-04-07 | 广州市进德生物科技有限公司 | Oxidized low-density lipoprotein protective agent, detection kit for determining oxidized low-density lipoprotein and application of detection kit |
| CN115932239B (en) * | 2022-12-22 | 2024-02-20 | 广州市进德生物科技有限公司 | Oxidized low-density lipoprotein protective agent, detection kit for determining oxidized low-density lipoprotein and application of detection kit |
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