CN2783324Y - Chip reagent box for cardiovascular diseases diagnosis and predicating multiple indes protein - Google Patents
Chip reagent box for cardiovascular diseases diagnosis and predicating multiple indes protein Download PDFInfo
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- CN2783324Y CN2783324Y CN 200520041211 CN200520041211U CN2783324Y CN 2783324 Y CN2783324 Y CN 2783324Y CN 200520041211 CN200520041211 CN 200520041211 CN 200520041211 U CN200520041211 U CN 200520041211U CN 2783324 Y CN2783324 Y CN 2783324Y
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Abstract
The utility model relates to integration detecting reaction plate for the diagnosis and the prediction of eight indexes of cardiovascular diseases and a protein chip kit. A reaction orifice plate comprises a basal plate and reaction holes arranged on the basal plate. The reaction holes comprise 12 to 200 sample holes and 5 to 8 standard holes, wherein the bottom of each of the reaction holes is provided with solid phase carriers and the solid phase carriers are coated with microarrays of eight antibodies, such as anti-crp (c-reactive protein), anti-Myoglobin (myoglobin), anti-cTnI (cardiac troponin I), anti-cTnT (cardiac troponin T), anti-nt-probnp (n-tail end brain natriuretic peptide), anti-ck-mb (creatine kinase isozymes-mb), anti-h-fabp (heart-type fatty acid binding protein), anti-GPBB (glycogen phosphorylase isoenzyme BB), etc. The reaction plate and the kit of the utility model can realize the diagnosis, the prediction, and the simultaneous detection of the eight indexes of the cardiovascular diseases for a plurality of people simply, rapidly and exactly.
Description
Technical field
The utility model belongs to external clinical examination and biochip technology field, exactly, is a kind of diagnosis that is suitable for angiocardiopathy, eight index (CRP of prediction and prognosis, Myoglobin, cTnI, cTnT, NT-proBNP, CK-MB, FABP and GPBB) reaction plate and the kit of integrated detection
Background technology
Angiocardiopathy comprises heart and vascular system disease.Since 20th century, along with the progress of civilization of human society, medical technology is developed significantly, and infectious diseases is progressively controlled.And expanding economy, the change of people's living standard and life style makes human diseases spectrum that marked change also take place, promptly turn to Chronic Non-Communicable Diseases be main to change.Wherein angiocardiopathy western countries progressively become cause dead main diseases because of, the death toll that is caused by angiocardiopathy accounts for 1/4 of the total death toll in the world, " the No.1 killer " who is called as human health, the serious threat mankind's life and quality of life cause the waste of huge manpower and economic resources.Therefore setting up a cover can be accurately, the detection system of prediction quickly and easily and diagnosis of cardiovascular diseases is extremely urgent.
Present modal angiocardiopathy is a coronary atherosclerotic heart disease.Coronary atherosclerotic heart disease abbreviates coronary heart disease as, is that the tube chamber that caused by coronary atherosclerosis blocks institute and causes, and further cause myocardial ischemia, the caused heart tissue necrosis of anoxic and the functional change of coronary artery.
A large amount of coronary atherosclerosiies that studies show that itself are the processes of an inflammation, and lasting low-level inflammation is indicating the possibility that angiocardiopathy takes place.In inflammation index, C-reactive protein (high sensitivity-CRP (hs-CRP)) is to cardiovascular at present, and the cerebrovascular and peripheral artery disease have best prediction and prognosis effect.
When coronary heart attack, the myocardial damage that hypoxic-ischemic caused of cardiac muscle can make some cardiac muscle cell's albumen be released into blood, and these indicator proteins have coronary heart disease diagnosis and important meaning.At present, it is CK-MB that coronary heart disease index the most commonly used detects in hospital, but its status is just replaced by " the golden index " that cTnI and these two of cTnT have high sensitivity and high specific simultaneously.But the time that these two indexs are released into blood is later relatively, therefore also needs Myoglobin, and early stage diagnosis is carried out in the cooperation of these three indexs of FABP and GPBB.
Except coronary heart disease, congestive heart failure also is a kind of common angiocardiopathy, and NT-proBNP is the sensitiveest and effective index of this disease of diagnosis.
Diagnosis and prediction to angiocardiopathy mainly is that single index is detected at present, wastes time and energy.The utility model is by realizing the generation and the development of angiocardiopathy are comprehensively predicted to the fast detecting of eight indexs, and diagnosis and prognosis have important directive significance to the prevention and the treatment of angiocardiopathy.
Summary of the invention
The purpose of this utility model provides a kind of can the prediction angiocardiopathy simultaneously, the method that diagnosis and prognosis are judged.Our kit passes through to detect the CRP in serum or the blood plasma, Myoglobin, and cTnI, cTnT, NT-proBNP, CK-MB, these eight indexs of FABP and GPBB reach this purpose.
In first aspect of the present utility model, a kind of diagnosis of angiocardiopathy is provided and has predicted eight index integral investigating reaction plates, the reaction orifice plate comprises substrate and the reacting hole that is positioned on the substrate, described reacting hole comprises 12-200 sample aperture, 5-8 standard items hole, wherein, it is characterized in that: solid phase carrier is arranged at the bottom of each reacting hole, and bag is by anti-CRP, anti-Myoglobin, anti-cTnI on solid phase carrier, anti-cTnT, anti-NT-proBNP, anti-CK-MB, the microarray of eight kinds of antibody such as anti-FABP and anti-GPBB.In each reacting hole, also be provided with a location reference point.
In a preference, described solid phase carrier is NC film or PDVF film.
In another preference, the quantity of described sample aperture is 4-195.
In another preference, described anti-CRP, anti-Myo, anti-cTnI, anti-cTnT, anti-NT-proBNP, anti-CK-MB, eight kinds of antibody such as anti-FABP and anti-GPBB respectively have 2 point of samples on solid phase carrier.
In another preference, the every hole of described reacting hole is provided with a positioning reference point.
In second aspect of the present utility model, a kind of diagnosis of angiocardiopathy is provided and has predicted eight integrated detection kit of index, it comprises the diagnosis of the angiocardiopathy that the utility model is above-mentioned and predicts six index integral investigating reaction plates.
In another preference, also comprise: enzyme mark working fluid, detection liquid and standard items.
In another preference, described enzyme mark working fluid is made up of the corresponding antigens or the monoclonal antibody of HRP mark, and described detection liquid is made up of shiner and two kinds of reagent of hydrogen peroxide, mixes before using.
In another preference, described shiner is luminol, different luminol or derivatives thereof.
Description of drawings
Fig. 1 is the point sample synoptic diagram of eight indexs that angiocardiopathy detects on a kind of solid phase carrier of the utility model
Fig. 2 is each antigen that carries out in the reacting hole of the present utility model---the synoptic diagram of antibody response
Embodiment
The key of eight integrated detections of index of angiocardiopathy is to make sensitivity that eight indexs detect and specificity all reach requirement guaranteeing the result accurately and reliably, and corresponding difficult point is how respectively the detect index of concentration difference up to 10000 times to be incorporated in the hole and to detect.The extensive studies through going deep into, the utility model adopts two kinds of different reactive modes to detect, and the detection reaction plate done improvement, the microarray detection technique of the utility model employing simultaneously, enzyme immunity indirect method and chemiluminescence detection technology, improve the sensitivity and the specificity that detect greatly, thereby solved the problems referred to above.
Particularly, for improving eight index (CRP of angiocardiopathy, Myoglobin, cTnI, cTnT, NT-proBNP, CK-MB, FABP and GPBB) detection efficiency, the utility model is used the microarray detection technique, desmoenzyme immunity indirect method and chemiluminescence detection technical design eight index integral investigating reaction plates of a kind of angiocardiopathy, the reaction orifice plate comprises substrate and the reacting hole that is positioned on the substrate, described reacting hole comprises 12-200 sample aperture, 5-8 standard items hole, wherein, it is characterized in that: on the base plane of each reacting hole solid phase carrier is arranged, and bag is by anti-CRP, anti-Myoglobin, anti-cTnI on solid phase carrier, anti-cTnT, anti-NT-proBNP, anti-CK-MB, the microarray of eight kinds of antibody such as anti-FABP and anti-GPBB.
The quantity of each reacting hole is not particularly limited, and sample aperture is 12-200 usually, preferably 5-50.Standard items hole 3-10, preferably 5-8.
In the utility model, the shape and the size of reacting hole are not particularly limited, and can be circular, square, sexangle, ellipse or other shapes.Size is generally diameter 0.5-5cm, and the degree of depth is 0.5-2cm.
In the utility model, the material of reaction plate substrate can be selected this area various materials commonly used for use, and preferable baseplate material comprises glass, plastics, pottery etc.
In the utility model, bag can be selected for use this area various solid phase carriers commonly used by the solid phase carrier of angiocardiopathy antibody, comprise (but being not limited to): glass sheet, plastics, cellulose nitrate (NC) film, PDVF film etc., preferable solid phase carrier comprises NC film, PDVF film.
On solid phase carrier, anti-CR is respectively arranged, anti-Myoglobin, anti-cTnI, anti-cTnT, anti-NT-proBNP, anti-CK-MB, eight kinds of antibody 1-3 point of samples such as anti-FABP and anti-GPBB, preferably 2 point of samples.In addition, preferably on solid phase carrier, also comprise an anchor point (see figure 1).Distance between each point of sample is generally 0.1-5mm.
On the other hand, the utility model also provides eight indexs of a kind of angiocardiopathy integrated detection protein chip kit, and this kit can be realized the parallelization of many indexs, microminiaturization, high throughput testing.It comprises reaction orifice plate of the present utility model, cleansing solution, enzyme mark working fluid, detects liquid (A, B), standard items etc.Wherein, enzyme mark working fluid is made up of the corresponding antigens or the monoclonal antibody of HRP mark, detects liquid and is made up of shiner and two kinds of reagent of hydrogen peroxide, and shiner is luminol, different luminol or derivatives thereof, mixes before using.
In the utility model, the detection of each index adopts enzyme to exempt from sandwich method or competition law, detects principle shown in Fig. 2 sandwich method and Fig. 3 competition law.Be described in detail as follows:
1) specific antibody is connected with solid phase carrier, forms insolubilized antibody.Unconjugated site is removed in sealing.
2) add and examined serum and enzyme mark solution, enzyme mark solution comprises eight corresponding enzyme labelled antibody (sandwich method) or enzyme-labelled antigens (competition law) that detect indexs.For the index that adopts sandwich method to detect, special enzyme labelled antibody (is illustrated as
(be illustrated as with the antigen in the serum
Form compound, and further (be illustrated as with insolubilized antibody
Form the double antibodies sandwich compound.For the index that adopts competition law to detect, special enzyme-labelled antigen (is illustrated as
(be illustrated as with the antigen in the serum
(be illustrated as with insolubilized antibody competitively
In conjunction with, form antigen antibody complex.
3) unconjugated antibody, antigen and impurity are removed in washing.
4) add substrate: the substrate for enzymatic activity on the solid phase, chemiluminescence, CCD detects, and the amount of being examined antigen in light intensity and the sample is proportionate (sandwich method) or negative correlation (competition law).
The utility model has fully utilized protein microarray detection technique dexterously, and enzyme immunity sandwich method and competition law, and chemiluminescence detection technology detect when having realized eight indexs of angiocardiopathy.
Below in conjunction with specific embodiment, further set forth the utility model.Should be understood that these embodiment only to be used to the utility model is described and be not used in the restriction scope of the present utility model.The experimental technique of unreceipted actual conditions in the following example, usually according to normal condition, people such as Sambrook for example, molecular cloning: laboratory manual (New York:Cold Spring HarborLaboratory Press, 1989) condition described in, or the condition of advising according to manufacturer.
Embodiment 1
The preparation of detection reaction plate
Get 48 orifice plates, 42 sample aperture are set, 6 standard items holes.Place a solid phase carrier (NC film or PDVF film) in the bottom of each reacting hole, and on solid phase carrier, wrap by anti-CRP, anti-Myoglobin, anti-cTnI, anti-cTnT, anti-NT-proBNP, anti-CK-MB, the little array of protein (Fig. 1) of eight kinds of antibody such as anti-FABP and anti-GPBB.
Embodiment 2
The kit preparation
Preparation contains the kit of eight integrated detections of index of an angiocardiopathy, and it contains one of the reaction orifice plate (24-48 person-portion) of embodiment 1,1 bottle of concentrated solution for washing, and 1 bottle of enzyme mark working fluid detects one bottle of liquid A, detects one bottle of liquid B, the standard items bottle.
Concentrated solution for washing, enzyme mark working fluid, detection liquid A, detection liquid B
Embodiment 3
Eight integrated detections of index of angiocardiopathy
By the following method each test blood please be detected by sample:
1. add detect blood please or standard items in the respective reaction hole, the 75ul/ hole.
2. enzyme-added mark working fluid, the 75ul/ hole.
3.37 ℃ insulation oscillating reactions 60 minutes.
4. with washing lotion washing 4-6 time, each 2 minutes, vibration.
5. add and detect liquid 20ul/ hole, make and detect the bottom that liquid is uniformly distributed in the hole.
6.CCD detect, look luminous signal intensity exposure 30-60 second.
The result judges:
According to the CutOff value of different indexs, normative reference product gradient curve, the yin and yang attribute of judgement sample serum.
Meanwhile, with conventional method the blood serum sample of testing is carried out conventional sense.The result shows that the test result that draws with kit of the present utility model is identical with conventional method.
All quote in this application as a reference at all documents that the utility model is mentioned, just quoted as a reference separately as each piece document.Should be understood that in addition those skilled in the art can make various changes or modifications the utility model after having read above-mentioned teachings of the present utility model, these equivalent form of values fall within the appended claims of the application institute restricted portion equally.
Claims (9)
1. the diagnosis of an angiocardiopathy and predict eight index integral investigating reaction plates is characterized in that, the reaction orifice plate comprises substrate and is positioned at reacting hole on the substrate, described reacting hole comprises 12-200 sample aperture, 5-8 standard items hole, wherein, it is characterized in that: solid phase carrier is arranged at the bottom of each reacting hole, and bag is by anti-CRP, anti-Myo, anti-cTnI on solid phase carrier, anti-cTnT, anti-NT-proBNP, anti-CK-MB, the microarray of eight kinds of antibody such as anti-FABP and anti-GPBB.In each reacting hole, also be provided with a location reference point.
2. reaction plate as claimed in claim 1 is characterized in that, described solid phase carrier is NC film or PDVF film.
3. reaction plate as claimed in claim 1 is characterized in that, the quantity of described sample aperture is 4-195.
4. reaction plate as claimed in claim 1 is characterized in that, described anti-CRP, and anti-CRP, anti-Myo, anti-cTnI, anti-cTnT, anti-NT-proBNP, anti-CK-MB, eight kinds of antibody such as anti-FABP and anti-GPBB respectively have 2 point of samples on solid phase carrier.
5. reaction plate as claimed in claim 1 is characterized in that, also is provided with a location reference point in each reacting hole.
6. the diagnosis of an angiocardiopathy and predict eight integrated detection protein chip kits of index is characterized in that it comprises the diagnosis of the described angiocardiopathy of claim 1 and predicts eight index integral investigating reaction plates.
7. kit as claimed in claim 6 is characterized in that, also comprises: enzyme mark working fluid, detect liquid and standard items.
8. kit as claimed in claim 7 is characterized in that, described enzyme mark working fluid is made up of the corresponding antigens or the monoclonal antibody of HRP mark, and described detection liquid is made up of shiner and two kinds of reagent of hydrogen peroxide, mixes before using.
9. kit as claimed in claim 8 is characterized in that, described shiner is luminol, different luminol or derivatives thereof.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200520041211 CN2783324Y (en) | 2005-04-28 | 2005-04-28 | Chip reagent box for cardiovascular diseases diagnosis and predicating multiple indes protein |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200520041211 CN2783324Y (en) | 2005-04-28 | 2005-04-28 | Chip reagent box for cardiovascular diseases diagnosis and predicating multiple indes protein |
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| Publication Number | Publication Date |
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| CN2783324Y true CN2783324Y (en) | 2006-05-24 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN 200520041211 Expired - Lifetime CN2783324Y (en) | 2005-04-28 | 2005-04-28 | Chip reagent box for cardiovascular diseases diagnosis and predicating multiple indes protein |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EA013943B1 (en) * | 2010-01-11 | 2010-08-30 | Общество С Ограниченной Ответственностью Научно-Производственное Объединение "Биотест" (Ооо Нпо "Биотест") | The assay kit “cardio test” for immunochromatographic detection of heart fatty acid binding protein and troponin i in whole blood samples for express diagnosis of myocardial infarction |
| CN101819205A (en) * | 2010-06-03 | 2010-09-01 | 中国人民解放军第三军医大学 | Human N-terminal pro-B-type natriuretic peptide (NT-proBNP) immunoassay kit and preparation method thereof |
| WO2011068680A1 (en) * | 2009-12-03 | 2011-06-09 | Abbott Laboratories | Assay for diagnosis of cardiac myocyte damage |
| CN103959060A (en) * | 2011-09-30 | 2014-07-30 | 私募蛋白质体公司 | Cardiovascular risk event prediction and uses thereof |
| US8835120B2 (en) | 2009-12-02 | 2014-09-16 | Abbott Laboratories | Assay for cardiac troponin-T (cTnT) |
| CN106290883A (en) * | 2015-06-01 | 2017-01-04 | 上海凯创生物技术有限公司 | GPBB colloidal gold method detection kit |
| CN107515307A (en) * | 2017-10-13 | 2017-12-26 | 基蛋生物科技股份有限公司 | A kind of immune cardiovascular eight Indexs measure test strips of dry type |
| CN108627650A (en) * | 2018-03-23 | 2018-10-09 | 南京秋浦生物科技有限公司 | The preparation method and application of the protein chip of marker detection is infected for pet |
| CN113533715A (en) * | 2020-04-22 | 2021-10-22 | 武汉市朗典精医生物科技有限公司 | Preparation of double-index immunochromatography kit for detecting hFABP/GPBB |
-
2005
- 2005-04-28 CN CN 200520041211 patent/CN2783324Y/en not_active Expired - Lifetime
Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8652788B2 (en) | 2009-12-02 | 2014-02-18 | Abbott Laboratories | Assay for diagnosis of cardiac myocyte damage |
| US8835120B2 (en) | 2009-12-02 | 2014-09-16 | Abbott Laboratories | Assay for cardiac troponin-T (cTnT) |
| CN102725633B (en) * | 2009-12-03 | 2016-02-03 | 雅培制药有限公司 | The mensuration damaged for diagnosing cardiac muscle cell |
| CN102725633A (en) * | 2009-12-03 | 2012-10-10 | 雅培制药有限公司 | Assay for diagnosis of cardiac myocyte damage |
| JP2013513108A (en) * | 2009-12-03 | 2013-04-18 | アボット・ラボラトリーズ | Assays for the diagnosis of cardiomyocyte injury |
| WO2011068680A1 (en) * | 2009-12-03 | 2011-06-09 | Abbott Laboratories | Assay for diagnosis of cardiac myocyte damage |
| JP2015180888A (en) * | 2009-12-03 | 2015-10-15 | アボット・ラボラトリーズＡｂｂｏｔｔ Ｌａｂｏｒａｔｏｒｉｅｓ | Assay for diagnosis of cardiac myocyte damage |
| EA013943B1 (en) * | 2010-01-11 | 2010-08-30 | Общество С Ограниченной Ответственностью Научно-Производственное Объединение "Биотест" (Ооо Нпо "Биотест") | The assay kit “cardio test” for immunochromatographic detection of heart fatty acid binding protein and troponin i in whole blood samples for express diagnosis of myocardial infarction |
| CN101819205A (en) * | 2010-06-03 | 2010-09-01 | 中国人民解放军第三军医大学 | Human N-terminal pro-B-type natriuretic peptide (NT-proBNP) immunoassay kit and preparation method thereof |
| CN103959060A (en) * | 2011-09-30 | 2014-07-30 | 私募蛋白质体公司 | Cardiovascular risk event prediction and uses thereof |
| CN106290883A (en) * | 2015-06-01 | 2017-01-04 | 上海凯创生物技术有限公司 | GPBB colloidal gold method detection kit |
| CN106290883B (en) * | 2015-06-01 | 2018-06-29 | 上海凯创生物技术有限公司 | Glycogen phosphorylase isoenzyme BB colloidal gold method detection kit |
| CN107515307A (en) * | 2017-10-13 | 2017-12-26 | 基蛋生物科技股份有限公司 | A kind of immune cardiovascular eight Indexs measure test strips of dry type |
| CN108627650A (en) * | 2018-03-23 | 2018-10-09 | 南京秋浦生物科技有限公司 | The preparation method and application of the protein chip of marker detection is infected for pet |
| CN113533715A (en) * | 2020-04-22 | 2021-10-22 | 武汉市朗典精医生物科技有限公司 | Preparation of double-index immunochromatography kit for detecting hFABP/GPBB |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Assignee: Shanghai Yulong Biological Science And Technology Co., Ltd. Assignor: Mu Haidong Contract fulfillment period: 2008.8.8 to 2013.8.8 Contract record no.: 2008310000195 Denomination of utility model: Multi-index protein chip inspection reagent unit of cardiovascular disease diagnosis and prediction Granted publication date: 20060524 License type: Exclusive license Record date: 20081021 |
|
| LIC | Patent licence contract for exploitation submitted for record |
Free format text: EXCLUSIVE LICENSE; TIME LIMIT OF IMPLEMENTING CONTACT: 2008.8.8 TO 2013.8.8; CHANGE OF CONTRACT Name of requester: SHANGHAI YULONG BIOLOGICAL TECHNOLOGY CO., LTD. Effective date: 20081021 |
|
| EC01 | Cancellation of recordation of patent licensing contract |
Assignee: Shanghai Yulong Biological Science And Technology Co., Ltd. Assignor: Mu Haidong Contract record no.: 2008310000195 Date of cancellation: 20130823 |
|
| LICC | Enforcement, change and cancellation of record of contracts on the licence for exploitation of a patent or utility model | ||
| EE01 | Entry into force of recordation of patent licensing contract | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Assignee: Shanghai Yulong Biological Science And Technology Co., Ltd. Assignor: Mu Haidong Contract record no.: 2014310000011 Denomination of utility model: Multi-index protein chip inspection reagent unit of cardiovascular disease diagnosis and prediction Granted publication date: 20060524 License type: Exclusive License Record date: 20140121 |
|
| C17 | Cessation of patent right | ||
| CX01 | Expiry of patent term |
Expiration termination date: 20150428 Granted publication date: 20060524 |