CN106226521A - A kind of diagnostic kit detecting Lp PLA2 - Google Patents
A kind of diagnostic kit detecting Lp PLA2 Download PDFInfo
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- CN106226521A CN106226521A CN201610611222.6A CN201610611222A CN106226521A CN 106226521 A CN106226521 A CN 106226521A CN 201610611222 A CN201610611222 A CN 201610611222A CN 106226521 A CN106226521 A CN 106226521A
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- adjuvant
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- pla2
- buffer
- diagnostic kit
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/916—Hydrolases (3) acting on ester bonds (3.1), e.g. phosphatases (3.1.3), phospholipases C or phospholipases D (3.1.4)
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/28—Neurological disorders
- G01N2800/2871—Cerebrovascular disorders, e.g. stroke, cerebral infarct, cerebral haemorrhage, transient ischemic event
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/32—Cardiovascular disorders
- G01N2800/324—Coronary artery diseases, e.g. angina pectoris, myocardial infarction
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- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The present invention relates to a kind of diagnostic kit detecting Lp PLA2, this test kit comprises medicament and adjuvant, wherein said medicament is formed by being coated buffer, lavation buffer solution, chromosomes of cell nuclei and Lp PLA2 antibody, described adjuvant is made up of normal human serum, urea peroxide, enzyme stabilizers and biological preservative, described medicament and the ratio of the volume of adjuvant are 3: 5 10, wherein being coated buffer is the pH carbonate buffer solution of 0.08M, (NH of 0.15M when lavation buffer solution is pH7.4 when being 9.61)2SO1.Cardiovascular disease is prevented to have important clinical meaning and economic results in society for the state of an illness.
Description
Technical field
The invention belongs to chemiluminescent polypeptide and field of immunology, be specifically related to a kind of according to Lp-PLA2 quantitative determination reagent kit
Background technology
The pathologic basis of cardiovascular and cerebrovascular disease is atherosclerosis.In recent years, research finds that atherosclerosis is a kind of scorching
Property reactive disorder, the product of inflammatory cell and release thereof is considered as the factor of topmost pro-atherogenic.At medicated porridge
Sample speckle is formed, progress and all have the participation of inflammatory reaction medium during finally rupturing.Platelet-activating factor acetylhydro-lase by
PLA2G7 gene code, the platelet-activating factor acetylhydrolase that is otherwise known as (PAF-AH), belong to PLA2 in phospholipase family
One, be serine rely on phospholipase, its catalysis activity need not Ca2+.Dividing of human plasma platelet-activating factor acetylhydro-lase
Son amount is 45KDa, and it is mainly generated by secretions such as macrophage, mononuclear cell, T lymphocyte, mastocyte, hepatocyte, and
Regulated by inflammatory mediator.Such as, gamma interferon and lipopolysaccharide suppress it to secrete, and platelet activating factor promotes that it is secreted.
The Main Function of platelet-activating factor acetylhydro-lase includes: produces dodecane acids Inflammatory substances, participate in phospholipid reconstruction and biomembrane
Stable equilibrium, lipoprotein metabolism, cell signal transmission, host response, the autologous disappearance of promotion body slough.
Acute cardiovascular and cerebrovascular disease is the most common type that atherosclerosis causes organ lesion.In recent years, along with right
Deepening continuously of atherosclerosis study, many inflammatory factors are found, and it is that atherosclerosis is sent out that inflammatory has been recognized
Central factor during exhibition.Traditional Inflammatory Mediators such as interleukin, tumor necrosis factor, intercellular adhesion molecule,
Vascular cell adhesion molecule, Fibrinogen, c reactive protein etc., promote atheromatous plaque formed in effect by
Gradually confirmed by numerous experiments.Platelet-activating factor acetylhydro-lase, as a kind of inflammatory reaction medium recently studied, has promotion
Atherosclerosis, has substantial connection with cardiocerebrovasculaevents events, and the diagnosis for cardiovascular and cerebrovascular disease adds one
Plant new means, and become the potential target target spot that such disease for the treatment of is new.
When atherosclerotic inflammatory development to the order of severity, Lp-PLA2 will be released in blood in a large number, causes it
Level in blood significantly increases, and therefore Lp-PLA2 concentration in blood can reflect the degree of inflammation of atherosclerotic plaque.
Therefore, by the Lp-PLA2 in detection blood, can effectively understand the degree of inflammation of atheromatous plaque and stablize
Property.Thus can early warning myocardial infarction and the generation of cerebral thrombosis, to prevention cardiovascular and cerebrovascular vessel accident there is considerable meaning
Justice.
Summary of the invention
In order to solve above-mentioned technical problem, the present invention provides a kind of diagnostic kit detecting Lp-PLA2, it is characterised in that should
Test kit comprises medicament and adjuvant, wherein said medicament by be coated buffer, lavation buffer solution, chromosomes of cell nuclei and
Lp-PLA2 antibody forms, and described adjuvant is made up of normal human serum, urea peroxide, enzyme stabilizers and biological preservative,
Described medicament and the ratio of the volume of adjuvant are 3: 5-10.
Described it is coated the carbonate buffer solution of 0.08M, 0.15M when dilution buffer is pH7.4 when buffer preferably pH is 9.6
(NH4)2SO4。
Described nuclei dyeing toner be preferably TMB and or bromination the third pyridine, further preferably by weight
TMB than 2: 1 and bromination the third pyridine composition.
Be coated buffer in described medicament, the volume ratio of dilution buffer is preferably 1: 1, and wherein chromosomes of cell nuclei accounts for described medicine
The 15-35% of agent cumulative volume.
In described adjuvant, normal human serum is preferably 5 with the volume ratio of urea peroxide: 2-3, and biological preservative and enzyme are steady
Determine the preferred 0.3g/ml of ratio of agent.
Described enzyme stabilizers is preferably made up of EDTA, sucrose, glycerol, NaCl, bovine serum albumin, Hydrazoic acid,sodium salt and water, further
Preferably by percentage by weight 0.1%EDTA, the sucrose of 3%, 52% glycerol, 1.3%NaCl, 1% bovine serum albumin and 0.02%
Hydrazoic acid,sodium salt and the water composition of 42.58%.
Described biological preservative is preferably the alcoholic solution of 50wt% epsilon-polylysine.
Beneficial effect
Can learn according to the narration of such scheme, coronary heart disease and palsy risk diagnosis reagent kit utilize special and High sensitivity to exempt from
Epidemic disease detection method (ELISA) carrys out the content of the Lp-PLA2 in detection by quantitative human serum or blood plasma, and then is used for predicting coronary heart disease, lacks
The danger of courageous and upright myocardial infarction and apoplexy because of.Prediction, prevention for the state of an illness have important clinical meaning and economic results in society.
The Lp-PLA2 antibody prepared by the present invention can high special Lp-PLA2 in blood sample be combined.The present invention's
People's Lp-PLA2 external diagnosis reagent case can monitor degree of inflammation and the stability thereof of atheromatous plaque effectively, can
Early warning myocardial infarction and the generation of cerebral thrombosis, prevent cardiovascular and cerebrovascular vessel accident.
The Lp-PLA2 quantitative determination reagent kit of the present invention is the Lp-PLA2 of 90-897ng/ml for sample concentration, the response rate)
97%.To low concentration (200ng/ml), middle concentration (500ng/ml) and high concentration (780ng/ml), in its group, the coefficient of variation is respectively
It is 3.7%, 5.0% and 4.1% (N=30), and the coefficient of variation between group is respectively 5.5%, 7.4% and 7.8% (N=10)；
Test kit True Positive Rate: 95.2%；Specificity (true negative rate): about 99%；False positive rate: 3%；False negative rate: 16.0%.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is further described in detail.
Lp-PLA2 antibody used in embodiment is purchased from Beijing Suo Laibao Science and Technology Ltd., and its character is as follows:
Target spot: 25；Concentration is: 1mg/ml；Antibody sources: Mus；Storage temperature :-20 DEG C；Hypotype: IgG.
In embodiment enzyme stabilizers be weight percentage 0.1%EDTA, the sucrose of 3%, 52% glycerol, 1.3%NaCl, 1% N
Serum albumin and 0.02% Hydrazoic acid,sodium salt and the water composition of 42.58%；Biological preservative is the ethanol of 50wt% epsilon-polylysine
Solution,
Embodiment 1
A kind of diagnostic kit detecting Lp-PLA2, this test kit comprises medicament and adjuvant, and wherein said medicament is by being coated
Buffer, lavation buffer solution, TMB and Lp-PLA2 antibody composition, described adjuvant is by just
Ordinary person's serum, urea peroxide, enzyme stabilizers and biological preservative composition, described medicament and the ratio of the volume of adjuvant are 3:
10, being wherein coated buffer is the pH carbonate buffer solution of 0.08M, 0.15M when lavation buffer solution is pH7.4 when being 9.6
(NH4)2SO4, TMB is subacidity.Its Chinese medicine is coated the volume of buffer, lavation buffer solution
Ratio is 1: 1, and wherein TMB accounts for the 30% of described medicament cumulative volume.Normal person in described adjuvant
Serum is 5: 3 with the volume ratio of urea peroxide, and the ratio of biological preservative and enzyme stabilizers is 0.3g/ml.
Embodiment 2
A kind of diagnostic kit detecting Lp-PLA2, this test kit comprises medicament and adjuvant, and wherein said medicament is by being coated
Buffer, lavation buffer solution, bromination the third pyridine and Lp-PLA2 antibody composition, described adjuvant is by normal human serum, peroxidating
Urea, enzyme stabilizers and biological preservative composition, described medicament and the ratio of the volume of adjuvant are 3: 10, are wherein coated buffer
The carbonate buffer solution of 0.08M when being 9.6 for pH, (the NH of 0.15M when lavation buffer solution is pH7.44)2SO4.In its Chinese medicine
Be coated buffer, the volume ratio of lavation buffer solution is 1: 1, and wherein bromination the third pyridine accounts for the 30% of described medicament cumulative volume.Described
In adjuvant, normal human serum is 5: 3 with the volume ratio of urea peroxide, and the ratio of biological preservative and enzyme stabilizers is 0.3g/
ml。
Embodiment 3
A kind of diagnostic kit detecting Lp-PLA2, this test kit comprises medicament and adjuvant, and wherein said medicament is by being coated
Buffer, lavation buffer solution, TMB, bromination the third pyridine and Lp-PLA2 antibody composition, described auxiliary
Agent is made up of normal human serum, urea peroxide, enzyme stabilizers and biological preservative, described medicament and the volume of adjuvant it
Ratio is 3: 10, and being wherein coated buffer is pH carbonate buffer solution of 0.08M when being 9.6, when lavation buffer solution is pH7.4
(the NH of 0.15M4)2SO4.Be coated buffer in its Chinese medicine, the volume ratio of lavation buffer solution is 1: 1, wherein 3,3 ', 5,5 '-four
Methyl biphenyl amine is 1: 1 with the weight ratio of bromination the third pyridine, and both account for the 30% of described medicament cumulative volume.In described adjuvant just
Ordinary person's serum is 5: 3 with the volume ratio of urea peroxide, and the ratio of biological preservative and enzyme stabilizers is 0.3g/ml.
The detailed description of the invention of the present invention is only the preferred embodiment of this creation, not in order to limit this creation, all in this creation
Spirit and principle within any modification, equivalent substitution and improvement etc. done, should be included in this creation protection domain it
In.
Claims (8)
1. the diagnostic kit detecting Lp-PLA2, it is characterised in that this test kit comprises medicament and adjuvant, Qi Zhongsuo
The medicament stated is formed by being coated buffer, lavation buffer solution, chromosomes of cell nuclei and Lp-PLA2 antibody, and described adjuvant is
Being made up of normal human serum, urea peroxide, enzyme stabilizers and biological preservative, described medicament and the ratio of the volume of adjuvant are
3∶5-10。
2. diagnostic kit as claimed in claim 1, it is characterised in that being coated buffer described in: is pH 0.08M when being 9.6
Carbonate buffer solution, (the NH of 0.15M when dilution buffer is pH7.44)2SO4。
3. diagnostic kit as claimed in claim 1, it is characterised in that: described nuclei dyeing toner is 3,3 ', 5,5 '-tetramethyl
Base benzidine and or bromination the third pyridine.
4. diagnostic kit as claimed in claim 3, it is characterised in that: described nuclei dyeing toner by the 3 of weight ratio 2: 1,
3 ', 5,5 '-tetramethyl benzidine and bromination the third pyridine composition.
5. diagnostic kit as claimed in claim 1, it is characterised in that: further, its Chinese medicine is coated buffer, dilute
The volume ratio releasing buffer is 1: 1, and wherein chromosomes of cell nuclei accounts for the 15-35% of described medicament cumulative volume.Described adjuvant
Middle normal human serum is 5: 2-3 with the volume ratio of urea peroxide, and the ratio of biological preservative and enzyme stabilizers is 0.3g/ml.
Measure test kit the most as claimed in claim 1, it is characterised in that: enzyme stabilizers is by EDTA, sucrose, glycerol, NaCl, cattle
Serum albumin, Hydrazoic acid,sodium salt and water composition.
Measure test kit the most as claimed in claim 1, it is characterised in that: enzyme stabilizers by percentage by weight 0.1%EDTA,
Sucrose, 52% glycerol, 1.3%NaCl, 1% bovine serum albumin and 0.02% Hydrazoic acid,sodium salt of 3% and the water composition of 42.58%.
Measure test kit the most as claimed in claim 1, it is characterised in that: described biological preservative is 50wt% epsilon-polylysine
Alcoholic solution.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610611222.6A CN106226521B (en) | 2016-07-25 | 2016-07-25 | A kind of diagnostic kit of detection Lp-PLA2 |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610611222.6A CN106226521B (en) | 2016-07-25 | 2016-07-25 | A kind of diagnostic kit of detection Lp-PLA2 |
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| Publication Number | Publication Date |
|---|---|
| CN106226521A true CN106226521A (en) | 2016-12-14 |
| CN106226521B CN106226521B (en) | 2018-11-06 |
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| CN201610611222.6A Active CN106226521B (en) | 2016-07-25 | 2016-07-25 | A kind of diagnostic kit of detection Lp-PLA2 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108844950A (en) * | 2018-04-25 | 2018-11-20 | 北京普赞生物技术有限公司 | A kind of disease meat quick testing reagent box and its detection method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1507565A (en) * | 2001-05-10 | 2004-06-23 | ҽ�����ϵͳ�������ι�˾ | Quantitative single-step immunoassay in lyophilised form |
| CN104730241A (en) * | 2015-03-02 | 2015-06-24 | 深圳市第二人民医院 | Kit used for detecting enzymatic activity of cardiovascnlar and cerebrovascular diseases |
-
2016
- 2016-07-25 CN CN201610611222.6A patent/CN106226521B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1507565A (en) * | 2001-05-10 | 2004-06-23 | ҽ�����ϵͳ�������ι�˾ | Quantitative single-step immunoassay in lyophilised form |
| CN104730241A (en) * | 2015-03-02 | 2015-06-24 | 深圳市第二人民医院 | Kit used for detecting enzymatic activity of cardiovascnlar and cerebrovascular diseases |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108844950A (en) * | 2018-04-25 | 2018-11-20 | 北京普赞生物技术有限公司 | A kind of disease meat quick testing reagent box and its detection method |
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| CN106226521B (en) | 2018-11-06 |
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