CN108918883A - A kind of immunofluorescent reagent box of rapid quantitative detection cardiac muscle troponin I, creatine kinase isozyme and myoglobins - Google Patents
A kind of immunofluorescent reagent box of rapid quantitative detection cardiac muscle troponin I, creatine kinase isozyme and myoglobins Download PDFInfo
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Abstract
The present invention provides the immunofluorescent reagent box of a kind of rapid quantitative detection cardiac muscle troponin I, creatine kinase isozyme and myoglobins, is related to field of biotechnology.The kit includes Immunofluorescence test paper strip, and test strips include support pad, detection film, conjugates gasket, filtering gasket, water suction gasket and sample gasket；Sample gasket is coated with red blood cell blocking agent, it detects film and is equipped with cardiac muscle troponin I, creatine kinase isozyme and myoglobins calibration tape and quality control band, cardiac muscle troponin I, creatine kinase isozyme and myoglobins calibration tape are coated with cardiac muscle troponin I antibody A, creatine kinase isozyme antibody A and myoglobins antibody A respectively, it is coated with anti-Streptavidin antibody on quality control band, heterophile antibody blocking agent, fluorescent dye conjugates are coated on conjugates gasket.Cardiac muscle troponin I, creatine kinase isozyme and myoglobins while immunofluorescent reagent box of the present invention can be quick, accurate, highly sensitive in detection sample, facilitate the Rapid&Early diagnosis of acute myocardial infarction AMI.
Description
Technical field
The present invention relates to field of biotechnology, and in particular to a kind of rapid quantitative detection cardiac muscle troponin I, creatine kinase
The immunofluorescent reagent box of isodynamic enzyme and myoglobins.
Background technique
Acute myocardial infarction AMI (AMI) is that coronary artery is acute, myocardial necrosis caused by duration hypoxic-ischemic.Clinically
There is violent and lasting retrosternal pain, rest and nitrate esters medicine are unable to complete incidence graph, with serum enzyme activities more
Increase and progressive ECG Change, can complicated by arrhythmia, shock or heart failure, often can threat to life.Acute myocardial stalk
It is dead occur after, if cannot be clarified a diagnosis within a few hours and suitably be treated, cardiac muscle will be because irreversible caused by ischemic, anoxic
Necrosis；During the diagnosis of acute myocardial infarction AMI, electrocardiogram is one of indispensable detection project, but a large amount of clinical datas
Show that its sensibility only has 50%, therefore, the joint-detection of serum cardiac damage markers becomes current acute myocardial infarction AMI and examines
Disconnected main trend.
Cardiac muscle troponin I (cTnI)：Troponin is contraction of muscle regulatory protein, including the different Asia of three structures
Base, i.e. troponin T (TnT), Troponin I (TnI) and troponin C (TnC).Cardiac muscle troponin I is a kind of structure egg
It is white, it is combined when actomyosin is in stationary state, inhibits the atpase activity of actomyosin.I subunit structure in cardiac muscle
(cTnI) it is different from other musculatures, there is Cardiac-specific；Cardiac muscle troponin I molecular weight is small simultaneously, after myocardial damage,
It discharges rapidly into blood, blood level increases after 4-6 hours, and can maintain 4-10 days.Due to the Cardiac-specific of its height
And sensitivity, Troponin I have become current optimal myocardial infarction marker.Global myocardial infarction working group, the European heart
Popular name for association (ESC), American Society of Cardiology (ACC), American Heart Association (AHA), European Society of Hypertension (EHS) and generation
Boundary's health organization (WHO), " goldstandard ", the acute coronary that cardiac muscle troponin I is set to acute myocardial infarction AMI (AMI) diagnosis are comprehensive
The preferred marker of simulator sickness (ACS) risk stratification.
Creatine kinase isozyme (CK-MB)：Creatine kinase (CK) isodynamic enzyme is by M subunit and B subunit group in different ways
The dimer of synthesis, is primarily present skeletal muscle and cardiac muscle cell, and there are three types of isodynamic enzymes：CK-MM, CK-MB, CK-BB, wherein flesh
Activity of the acid kinase isodynamic enzyme in cardiac muscle is 5 times in skeletal muscle, and creatine kinase isozyme is after acute myocardial infarction
It increases within 4-8 hours, peaks within 12-20 hours, it is more excellent for the specificity and sensibility of Diagnosis of Acute Myocardial Infarction.
Myoglobins (Myoglobin)：Myoglobins is the chromoprotein being present in cardiac muscle and skeletal muscle endochylema, in urgency
Property myocardial infarction (AMI) after the onset of raisings can be detected within 1-4 hour in blood, reach within 6-7 hours peak value, be to represent myocardium damage
One of earliest period biochemical marker of wound, the sensibility with height.But myoglobins is without Cardiac-specific, with myocardium myo calcium
Protein I and creatine kinase isozyme joint-detection can help to the early diagnosis of acute myocardial infarction AMI.
In the prior art, it is same to lack the short cardiac muscle troponin I of accuracy height, high sensitivity, detection time, creatine kinase
The kit of work enzyme and myoglobins.
Summary of the invention
The object of the present invention is to provide a kind of rapid quantitative detection cardiac muscle troponin I, creatine kinase isozyme and flesh are red
The immunofluorescent reagent box of albumen, can quickly, it is accurate, highly sensitive while detection whole blood, serum or blood plasma in cardiac muscle
Troponin I, creatine kinase isozyme and myoglobins facilitate the Rapid&Early diagnosis of acute myocardial infarction AMI.
The purpose of the present invention adopts the following technical scheme that realization.
A kind of immunofluorescent reagent of rapid quantitative detection cardiac muscle troponin I, creatine kinase isozyme and myoglobins
Box, including immunofluorescent reagent item, the test strips include support pad, the detection film being set in the support pad, institute
The both ends for stating detection film are connected separately with filtering gasket and water suction gasket, and the filtering gasket is connected with conjugates gasket, described
The other end of conjugates gasket is connected with sample gasket；The sample gasket is coated with red blood cell blocking agent, extremely from sample gasket
It is red that cardiac muscle troponin I calibration tape, creatine kinase isozyme calibration tape, flesh are successively arranged on detection film described in water suction gasket direction
Albumen test band and quality control band, the cardiac muscle troponin I calibration tape are coated with cardiac muscle troponin I antibody A, and the creatine swashs
Enzyme isoenzyme calibration tape is coated with creatine kinase isozyme antibody A, and the myoglobins calibration tape is coated with myoglobins antibody
A is coated with anti-Streptavidin antibody on the quality control band, be coated on the conjugates gasket heterophile antibody blocking agent,
Cardiac muscle troponin I antibody B, the creatine kinase isozyme antibody B and myoglobins antibody B of fluorochrome label.
In the present invention, the anti-human cardiac troponin of mouse that the cardiac muscle troponin I antibody A is available from Hytest company
I monoclonal antibody, article No. 4TC2；The anti-human flesh of mouse that the creatine kinase isozyme antibody A is available from Biospacific company
Acid kinase isozyme antibody, article No.：A27020067P；The mouse that the myoglobins antibody A is available from Biospacific company is anti-
Human muscle hemoglobin antibody, article No.：A27340228P；The anti-Streptavidin antibody is purchased from Prospec company, article No.：ANT-
345。
In the present invention, the heterophile antibody blocking agent is purchased from Scantibodies Laboratory company, article No.：
3KG775。
In the present invention, the red blood cell blocking agent is mouse anti human red blood cell monoclonal antibody.
In the present invention, the cardiac muscle troponin I antibody B is the anti-human myocardium myo calcium egg of mouse of Biospacific company
White I monoclonal antibody, article No. A34600384P, the creatine kinase isozyme antibody B are available from Biospacific company
The anti-human creatine kinase isozyme antibody of mouse, article No.：A27080359P；The myoglobins antibody B is available from Fitzgerald public affairs
The anti-human myoglobins antibody of the mouse of department, article No.：10-M50C.
In the present invention, the sample gasket, conjugates gasket, filtering gasket, detection film and water suction gasket are all set in
It arranges in support pad and linearly.
In the present invention, the cardiac muscle troponin I calibration tape, creatine kinase isozyme calibration tape, myoglobins test
Band is parallel with quality control band.
Kit of the present invention can be used for the myocardium myo in quick, accurate, highly sensitive detection whole blood, serum and blood plasma
Calcium protein I, creatine kinase isozyme and myoglobins facilitate the Rapid&Early diagnosis of acute myocardial infarction AMI.Test paper of the present invention
Item is limited to 0.08ng/ml to the detection of cTnI in sample, and the range of linearity of detection is 0.1ng/ml~40ng/ml；To CK-MB
Detection be limited to 0.5ng/ml, the range of linearity of detection is 1ng/ml~50ng/ml；5ng/ is limited to the detection of Myoglobin
Ml, the range of linearity of detection are 15ng/ml~400ng/ml.In the linear range, dose-response curve related coefficient (r) is not
Lower than 0.990.In addition, the coefficient of variation of test strips repeatability testing result of the present invention is equal<10.0%, illustrate this reagent detection knot
Fruit is reproducible, and precision is high.It is comprehensive that the clinical meaning of immunofluorescent reagent box of the present invention is that (1) facilitates acute coronary
The early diagnosis of simulator sickness and risk stratification；(2) estimates treating myocardial ischemia damage area；(3) antidiastole of various pectoralgia reasons；
(4) left ventricular function is assessed；(5) evaluation of clinical of certain therapeutic agents；(6) more frequently being early detection cTnI or
CK-MB, especially joint-detection Myoglobin can help to be diagnosed to be myocardial infarction as early as possible and be conducive to start as early as possible to control
It treats；(7) various myocardial damages are diagnosed：Myocarditis, myocardium wound, openheart surgery, periprocedural cardiac complication, Severe sepsis blood
Left heart failure caused by disease and septicopyemia, the diseases such as congested cardiac insufficiency.
Detailed description of the invention
Fig. 1 is the structure diagram of test strips of the present invention, wherein 1- sample gasket, 2- conjugates gasket, and 3- filters gasket, 4-
Detect film, 5- support pad, 6- water suction gasket.
Specific embodiment
The immunofluorescence that embodiment 1 prepares quantitative detection cardiac muscle troponin I, creatine kinase isozyme and myoglobins is tried
Agent item
Such as Fig. 1, the Immunofluorescence test paper strip of quantitative detection cardiac muscle troponin I, creatine kinase isozyme and myoglobins,
The test strips include support pad 5, the detection film 4 being set in support pad, and the both ends for detecting film are connected separately with filter pad
Piece 3 and water suction gasket 6, filtering gasket are connected with conjugates gasket 2, and the other end of conjugates gasket is connected with sample gasket 1,
Middle sample gasket is coated with red blood cell blocking agent, is successively arranged myocardium myo calcium from sample gasket to water suction gasket angle detecting film
Protein I calibration tape, creatine kinase isozyme calibration tape, myoglobins calibration tape and quality control band, cardiac muscle troponin I calibration tape packet
There is cardiac muscle troponin I antibody, creatine kinase isozyme calibration tape is coated with creatine kinase isozyme antibody, myoglobins
Calibration tape is coated with myoglobins antibody, and anti-Streptavidin antibody is coated on quality control band, is coated on conjugates gasket different
The agent of preferendum antibody blocking, the cardiac muscle troponin I antibody of fluorochrome label, creatine kinase isozyme antibody and myoglobins are anti-
Body.
Sample gasket is prepared with the following method：Mouse anti human red blood cell monoclonal antibody is used and contains 0.5% (matter
Measure percentage concentration) bovine serum albumin(BSA) PBS buffer solution, be configured to the solution of final concentration of 1.35mg/ml, use even application
Method gasket is coated with, after coating 37 DEG C dry 24 hours, obtain sample gasket, save backup.
Conjugates gasket is prepared with the following method：Mouse monoclone antibody against human cardiac troponin I (is purchased from
Biospacific company, article No. A34600384P) it is marked using biotin, using marked by streptavidin fluorescent dye
DylightTM800NHS Ester (is purchased from Thermo Scientific company, article No.：46421), then by the two by strepto- parent
Molar ratio with element and biotin is 1：4 are mixed in the phosphate buffer of pH=8.4,25 DEG C of incubation 30min, the two bridging shape
At cardiac muscle troponin I antibody-biotin-Streptavidin-fluorescence-conjugated object (being abbreviated as conjugates A).To contain 0.5%
The phosphate buffer of bovine serum albumin(BSA) is solvent, prepares the conjugates solution A of 0.1mg/ml.By the same work of the anti-human creatine kinase of mouse
Enzyme antibody (is purchased from Biospacific company, article No.：A27080359P it) is marked using biotin, using Streptavidin
Mark fluorescent dyestuff DylightTM800NHS Ester (is purchased from Thermo Scientific company, article No. 46421), then will
The two is 1 by the molar ratio of Streptavidin and biotin：4 are mixed in the phosphate buffer of pH=8.4,25 DEG C of incubations
30min, the two bridging form creatine kinase isozyme antibody-biotin-Streptavidin-fluorescence-conjugated object and (are abbreviated as being coupled
Object B).Using the phosphate buffer containing 0.5% bovine serum albumin(BSA) as solvent, compound concentration is that the conjugates B of 0.2mg/ml is molten
Liquid.The anti-human myoglobins antibody of mouse (is purchased from Fitzgerald company, article No.：It 10-M50C) is marked, is adopted using biotin
With marked by streptavidin fluorescent dye DylightTM800 NHS Ester (are purchased from Thermo Scientific company, article No.
46421) it is 1 that the two, which then, is pressed the molar ratio of Streptavidin and biotin,：4 are mixed in the phosphate buffer of pH=8.4
In, 25 DEG C of incubation 30min, the two bridging forms myoglobins antibody-biotin-Streptavidin-fluorescence-conjugated object and (is abbreviated as
Conjugates C).Using the phosphate buffer containing 0.5% bovine serum albumin(BSA) as solvent, compound concentration is the conjugates of 0.1mg/ml
C solution.From sample gasket to water suction gasket direction, conjugates are successively used with the mode uniformly crossed in fiberglass packing on piece
A, conjugates B and conjugates C solution are coated with, and coating region is band-like and is parallel to each other, and then use the side of even application
Heterophile antibody blocking agent of the formula in fiberglass packing on piece closing 0.7mg/ml (is purchased from Scantibodies
Laboratory company, article No.：3KG775), vacuum is drained, and obtains conjugates gasket, spare.
Detection film is prepared with the following method：From sample gasket to water suction gasket direction (i.e. by the direction of sample flowing),
Cardiac muscle troponin I calibration tape, creatine kinase isozyme calibration tape, myoglobins test are successively coated on nitrocellulose filter
Band and quality control band.Mouse monoclone antibody against human cardiac troponin I (being purchased from Hytest company, article No. 4TC2) is slow with phosphoric acid
Fliud flushing is diluted to the solution of final concentration of 2mg/ml, is coated on nitrocellulose membrane using the mode uniformly crossed and forms myocardium myo
Calcium protein I calibration tape.The anti-human creatine kinase isozyme antibody of mouse (is purchased from Biospacific company, article No.：A27020067P)
It is diluted to the solution of final concentration of 0.5mg/ml with phosphate buffer, is coated on nitrocellulose filter using the mode uniformly crossed
Upper formation creatine kinase isozyme calibration tape.The anti-human myoglobins antibody of mouse (is purchased from Biospacific company, article No.：
A27340228P it) is diluted to the solution of final concentration of 0.65mg/ml with phosphate buffer, is coated on using the mode uniformly crossed
Myoglobins calibration tape is formed on nitrocellulose filter.The anti-Streptavidin antibody of mouse (is purchased from Prospec company, article No.：
ANT-345 the phosphate buffer containing 0.5% bovine serum albumin(BSA)) is added, is configured to the solution of final concentration of 0.015mg/ml,
It is coated on nitrocellulose membrane using the mode uniformly crossed and forms quality control band.
The material of support pad is polyvinyl chloride.
The material of water suction gasket is that glass fibre and velveteen mixing (are purchased from Ahlstrom company, article No.：320).
The material for filtering gasket is cotton fiber.
Sample gasket, conjugates gasket, filtering gasket, detection film and water suction gasket are all set in support pad and are in
Straight line arrangement.
Cardiac muscle troponin I calibration tape, creatine kinase isozyme calibration tape, myoglobins calibration tape and quality control band are flat
Row.
In support pad, by sample gasket, conjugates gasket, filtering gasket and detection film along sample flow direction according to
Secondary overlap joint, water absorption pad oblique cut grafting obtain Immunofluorescence test paper strip in the end of detection membrane sample flow direction.
The sensitivity and the detection range of linearity of the kit of the present invention of embodiment 2
The application method of Immunofluorescence test paper strip (preparation of embodiment 1) of the present invention：Take 60 μ l drop of clinical patient plasma sample
Be added in the sample pad on piece of Immunofluorescence test paper strip, under the conditions of 18 DEG C be incubated for 8min after, be placed in auspicious Lay TZ-301 type be immunized it is glimmering
It is detected in optical detector, conventionally, calculates and obtain cTnI, CK-MB and Myoglobin concentration, specifically lead to
The fluorescence intensity on detection calibration tape and quality control band is crossed, relative optical density is calculated, it is then bent by relative optical density-concentration standard
Line computation show that cTnI (cardiac muscle troponin I), CK-MB (creatine kinase isozyme) and Myoglobin (myoglobins) are dense
Degree.By clinical cTnI positive plasma sample doubling dilution, and use Beckman Kurt Enhanced Accu TnI reagent (state
Eat medicine supervise tool (into) word 2013 the 2401177th) detection cTnI concentration, as a result 0.06ng/ml, 0.3125ng/ml,
0.625ng/ml,2.5ng/ml,10ng/ml,40ng/ml.By clinical CK-MB positive plasma sample doubling dilution and use creatine
Kinase isozyme (CK-MB) detection kit (immunofluorescence technique) (Soviet Union's tool note standard 20152401210) definite value, result
0.78125ng/ml,1.5625ng/ml,3.125ng/ml,12.5ng/ml, 50ng/ml.By clinical Myoglobin positive blood
It starches sample doubling dilution and (Soviet Union's tool is infused quasi- using myoglobins (Myoglobin) detection kit (immunofluorescence technique)
20152401211) definite value, result 12.5ng/ml, 25ng/ml, 50ng/ml, 100ng/ml, 400ng/ml.Using this hair
Bright Immunofluorescence test paper strip detects each concentration clinic cTnI positive sample, clinic CK-MB positive sample and clinic Myoglobin sun
Property sample, each diluted concentration tests 3 times, finds out the mean value of testing result respectively.The concentration detected with existing detection method
Regression analysis is carried out to two groups of data using Immunofluorescence test paper strip testing result mean value of the present invention as dependent variable y for independent variable x,
Equation of linear regression (i.e. dose-response curve) is found out, the related coefficient of linear regression is calculated.As a result such as the following table 1, table 2 and table
3。
The existing detection method of table 1 is with test strips of the present invention to the testing result of cTnI
| Definite value result (ng/ml) of the existing detection method to cTnI | Test strips test result average value (ng/ml) of the present invention |
| 0.06 | 0.10 |
| 0.3125 | 0.29 |
| 0.625 | 0.58 |
| 2.5 | 2.71 |
| 10 | 11.54 |
| 40 | 39.63 |
| Linear regression related coefficient | 0.99911 |
The existing detection method of table 2 is with test strips of the present invention to the testing result of CK-MB
| Definite value result (ng/ml) of the existing detection method to CK-MB | Test strips test result average value (ng/ml) of the present invention |
| 0.78125 | 1.13 |
| 1.5625 | 2.08 |
| 3.125 | 4.22 |
| 12.5 | 13.69 |
| 50 | 48.74 |
| Linear regression related coefficient | 0.99965 |
The existing detection method of table 3 is with test strips of the present invention to the testing result of Myoglobin
As a result it is found that test strips of the present invention are 0.1ng/ml~40ng/ml to the cTnI range of linearity detected, to CK-MB
The range of linearity of detection is 1ng/ml~50ng/ml, and the range of linearity to Myoglobin detection is 15ng/ml~400ng/
Ml, in the linear range, dose-response curve related coefficient (r) are not less than 0.990.
Detection of the test strips of the present invention to cTnI, CK-MB and Myoglobin in plasma sample is detected using the above method
Sensitivity, as a result, it has been found that test strips of the present invention are limited to 0.08ng/ml to the detection of cTnI in plasma sample, the detection to CK-MB
It is limited to 0.5ng/ml, 5ng/ml is limited to the detection of Myoglobin.
The repeatability and precision of the kit of the present invention of embodiment 3
1 part of clinical sample is detected using test strips of the present invention (preparation of embodiment 1), each sample repeats 10 detections.Tool
Body detecting method：Take clinical patient sample (serum) 60ul that the sample pad on piece of test strips of the present invention is added, under the conditions of 18 DEG C
It after being incubated for 8min, is placed in auspicious Lay TZ-301 type Immunofluorescence test instrument and is detected, by detection calibration tape and quality control band
Fluorescence intensity, and cTnI, CK-MB and Myoglobin concentration are obtained according to preset calculation formula and conversion curve, to same
Pattern detection result carries out average, variance analysis, and calculates the coefficient of variation, as a result such as table 4.
Testing result of the test strips of the present invention of table 4 to cTnI, CK-MB and Myoglobin concentration in clinical sample
The above results are shown：The repeated testing result coefficient of variation is equal<6%, illustrate that this reagent testing result is reproducible,
Precision is high.
The interference free performance of the kit of the present invention of embodiment 4
In order to verify the accuracy of Immunofluorescence test paper strip (2# in table 5) testing result that in embodiment 1 prepared by method, with
And in the detection process, the Multiple Antibodies in the test strips will not interfere the detection of important test object cardiac muscle troponin I, into
Following experiment is gone.
1#, 3#, 4#, 5# test strips are prepared using method in embodiment 1, the difference is that changing myocardium myo on detection film
Myocardium myo calcium egg on the coated antibody and conjugates gasket of calcium protein I calibration tape (being abbreviated as cTnI calibration tape) in conjugates A
White I antibody.Specifically it is shown in Table 5.
CTnI antibody conjugates situation in each test strips of table 5
1 part of cTnI quality-control product is detected simultaneously using 1#, 2#, 3#, 4#, 5# test strips, is repeated 5 times detection.CTnI quality-control product
Contain 0.15ng/mL cardiac muscle troponin I, 0ng/mL creatine kinase isozyme and 0ng/mL myoglobins.Specific detection side
Method：It takes cTnI quality-control product 60ul that the sample pad on piece of test strips is added, after being incubated for 8min under the conditions of 18 DEG C, is placed in auspicious Lay TZ-
It is detected in 301 type Immunofluorescence test instrument, by the fluorescence intensity on detection calibration tape and quality control band, and according to preset
Calculation formula and conversion curve obtain cTnI concentration, survey result to Quality Control product examine and carry out average, variance, deviation, sensitivity point
The coefficient of variation is analysed and calculates, as a result such as table 6.
Testing result of each test strips of table 6 to cTnI concentration in quality-control product
As can be seen from Table 6,1#, 4#, 5# test strips testing result deviation are larger, the side of 2#, 3# test strips testing result
Difference, the coefficient of variation and deviation are smaller.
When in order to determine the detection of 2#, 3# test strips, creatine kinase isozyme and myoglobins will not interfere cardiac muscle in sample
The testing result of Troponin I has prepared 5 kinds of samples containing cardiac muscle troponin I, myoglobins and creatine kinase isozyme
Product, such as table 7.5 kinds of samples are detected respectively using 2#, 3# test strips, and each sample is repeated 3 times detection.Detect calibration tape and matter
The fluorescence intensity taken is controlled, fluorescence intensity/quality control band fluorescence intensity of calibration tape is calculated, wherein cardiac muscle troponin I is tested
Fluorescence intensity/quality control band fluorescence intensity of band is abbreviated as cTnI/C, fluorescence intensity/Quality Control of creatine kinase isozyme calibration tape
The fluorescence intensity of band is abbreviated as CK/C, and fluorescence intensity/quality control band fluorescence intensity of myoglobins calibration tape is abbreviated as MYO/C.
Testing result is shown in Table 8,9.
The content of 75 kinds of sample Myocardial Troponin Is of table, myoglobins, creatine kinase isozyme
| Sample number into spectrum | Cardiac muscle troponin I | Myoglobins | Creatine kinase isozyme |
| 1 | 0.15ng/mL | 0ng/mL | 0ng/mL |
| 2 | 0.15ng/mL | 70ng/mL | 0ng/mL |
| 3 | 0.15ng/mL | 400ng/mL | 0ng/mL |
| 4 | 0.15ng/mL | 0ng/mL | 6ng/mL |
| 5 | 0.15ng/mL | 0ng/mL | 50ng/mL |
Table 8 detects the result of 5 kinds of samples respectively using 2# test strips and data are analyzed
It can be seen that the increase with myoglobin concentration from detection data (table 8) of the 2# test strips to sample 1,2,3,
The average value of cTnI/C has almost no change；It can be seen that from detection data of the 2# test strips to sample 1,4,5 as creatine swashs
The average value of the increase of enzyme isoenzyme concentration, cTnI/C has almost no change.Therefore, test strips are respectively tested in kit of the present invention
Band does not interfere with cardiac muscle troponin I Concentration Testing, and interband interferes with each other cross reaction and can ignore.
Table 9 detects the result of 5 kinds of samples respectively using 3# test strips and data are analyzed
It can be seen that the increase with myoglobin concentration from detection data (table 9) of the 3# test strips to sample 1,2,3,
The average value of cTnI/C is gradually increased；It can be seen that from detection data of the 3# test strips to sample 1,4,5 as creatine kinase is same
The average value of the increase of work enzyme concentration, cTnI/C is gradually increased.Therefore, other calibration tapes of 3# test strips are to cardiac muscle troponin I
Concentration Testing interferes significantly with, interband to interfere with each other cross reaction larger.
Claims (7)
1. the immunofluorescent reagent box of a kind of rapid quantitative detection cardiac muscle troponin I, creatine kinase isozyme and myoglobins,
It is characterized by comprising Immunofluorescence test paper strip, the test strips include support pad, the inspection that is set in the support pad
Film is surveyed, the both ends of the detection film are connected separately with filtering gasket and water suction gasket, and the filtering gasket is connected with conjugates pad
The other end of piece, the conjugates gasket is connected with sample gasket；The sample gasket is coated with red blood cell blocking agent, the inspection
It surveys film and is equipped with cardiac muscle troponin I calibration tape, creatine kinase isozyme calibration tape, myoglobins calibration tape and quality control band, institute
It states cardiac muscle troponin I calibration tape and is coated with cardiac muscle troponin I antibody A, the creatine kinase isozyme calibration tape is coated with
Creatine kinase isozyme antibody A, the myoglobins calibration tape are coated with myoglobins antibody A, are coated on the quality control band
Anti- Streptavidin antibody is coated with the myocardium myo of heterophile antibody blocking agent, fluorochrome label on the conjugates gasket
Calcium protein I antibody B, creatine kinase isozyme antibody B and myoglobins antibody B.
2. rapid quantitative detection cardiac muscle troponin I, creatine kinase isozyme and myoglobins are exempted from according to claim 1
Epidemic disease Fluorescence kit, it is characterised in that the anti-human myocardium myo calcium of the mouse that the cardiac muscle troponin I antibody A is available from Hytest company
Protein I monoclonal antibody, article No. 4TC2；The mouse that the creatine kinase isozyme antibody A is available from Biospacific company is anti-human
Creatine kinase isozyme antibody, article No.：A27020067P；The myoglobins antibody A is available from the mouse of Biospacific company
Anti-human myoglobins antibody, article No.：A27340228P；The anti-Streptavidin antibody is purchased from Prospec company, article No.：
ANT-345。
3. rapid quantitative detection cardiac muscle troponin I, creatine kinase isozyme and myoglobins according to claim 1 or claim 2
Immunofluorescent reagent box, it is characterised in that the heterophile antibody blocking agent is public purchased from Scantibodies Laboratory
Department, article No.：3KG775.
4. rapid quantitative detection cardiac muscle troponin I, creatine kinase isozyme and myoglobins are exempted from according to claim 3
Epidemic disease Fluorescence kit, it is characterised in that the red blood cell blocking agent is mouse anti human red blood cell monoclonal antibody.
5. rapid quantitative detection cardiac muscle troponin I, creatine kinase isozyme and myoglobins are exempted from according to claim 4
Epidemic disease Fluorescence kit, it is characterised in that the cardiac muscle troponin I antibody B is the anti-human myocardium myo of mouse of Biospacific company
Calcium protein I monoclonal antibody, article No. A34600384P, the creatine kinase isozyme antibody B are available from Biospacific public affairs
The anti-human creatine kinase isozyme antibody of the mouse of department, article No.：A27080359P；The myoglobins antibody B is available from
The anti-human myoglobins antibody of the mouse of Fitzgerald company, article No.：10-M50C.
6. rapid quantitative detection cardiac muscle troponin I, creatine kinase isozyme and myoglobins are exempted from according to claim 5
Epidemic disease Fluorescence kit, it is characterised in that the sample gasket, conjugates gasket, filtering gasket, detection film and water suction gasket are all provided with
It is placed in support pad and linearly arranges.
7. rapid quantitative detection cardiac muscle troponin I, creatine kinase isozyme and myoglobins are exempted from according to claim 6
Epidemic disease Fluorescence kit, it is characterised in that the cardiac muscle troponin I calibration tape, creatine kinase isozyme calibration tape, myoglobins
Calibration tape is parallel with quality control band.
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Application publication date: 20181130 |