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Kit and method for joint detection of cardiac troponin I, creatine kinase isozyme and myoglobin

A technology of cardiac troponin and creatine kinase, which is applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems of incomplete reaction, short reaction time, poor precision, etc., to solve the problem of poor precision and sensitivity , the effect of improving stability

Inactive Publication Date: 2020-08-14
巴迪泰(广西)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because this method directly coats the fluorescently labeled TnI, CK-MB, Myoglobin antibody complex and quality control complex combined with the antigen on the binding pad of the reagent strip, when the sample flows through the binding pad, the The reaction time between the antigen and the coated fluorescently labeled TnI, CK-MB, Myoglobin antibody complexes and quality control complexes is short, and the reaction may be incomplete, resulting in poor precision

Method used

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  • Kit and method for joint detection of cardiac troponin I, creatine kinase isozyme and myoglobin
  • Kit and method for joint detection of cardiac troponin I, creatine kinase isozyme and myoglobin
  • Kit and method for joint detection of cardiac troponin I, creatine kinase isozyme and myoglobin

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Experimental program
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Effect test

Embodiment 1

[0043] The manufacturing process of embodiment 1 reagent strip

[0044] Purchase and screen high-quality monoclonal antibodies, conduct pairing experiments, and screen out capture antibodies and detection antibodies with high specificity and good affinity. Use the coating diluent to adjust the concentration of CKMB detection antibody to 1-5mg / mL, streptavidin 1-5mg / mL, Myoglobin antibody 0.01-3mg / mL, chicken IgY 0.01-3mg / mL.

[0045] Antibody coating: Prepare the nitrocellulose membrane and place it correctly on the film sticking equipment, paste the nitrocellulose membrane on the pad card, set the spraying volume of the spraying equipment to 1 μL / cm, and the spraying speed to 9cm / s, and the detection line Spray-spray on the nitrocellulose membrane in parallel with the quality control line for coating, the distance between the test line and the quality control line is 2-3mm, and then place it in an oven at 37°C for 30-120 minutes.

[0046] Reagent strip film sticking and cutt...

Embodiment 2

[0048] The preparation process of embodiment 2 reaction buffer solution and detection particles

[0049] Preparation of fluorescent antibody complexes: Prepare the screened CK-MB capture antibody, Myoglobin capture antibody, TnI capture antibody, and Anti-Chicken IgY with 1*PBS to 1-5mg / mL respectively. Add 2%-20% (volume) 1M sodium bicarbonate solution to the above antibodies, and then add 1:0.01-1:0.5 (mass) fluorescent dyes into the solution, wrap them to avoid light, and place them on the mixer Stir the reaction for ≥ 4 hours, and control the temperature < 40°C. Purify the labeled fluorescent antibody complexes with chromatographic columns filled with cross-linked dextran G-25, collect the first outflowing fluorescent antibody complexes, and use a UV spectrophotometer to detect the prepared fluorescent antibody complexes The concentration and complexation ratio of the substance should be adjusted, and stored in the dark at 2-8°C for later use.

[0050] Preparation of bio...

Embodiment 3

[0055] Embodiment 3 adds formula and manufactures ID chip

[0056] Take the reagent strips, reaction buffers, TnI, CK-MB, and Myoglobin calibrators that have been tested for intermediate products. Each calibrator has at least 5 concentrations, and each concentration is tested at least 5 times. Scan the area of ​​each concentration calibrators The average value and concentration of the graph are used as a standard curve to obtain a linear equation, and the value of the equation is imported into the special software, and the blank original chip is connected to the computer, and the numerical code can be imported into the chip through the special software, and the chip label is affixed.

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Abstract

The invention discloses a kit and a method for joint detection of cardiac troponin I, creatine kinase isozyme and myoglobin. The kit contains: detection particles containing a fluorescently labeled TnI antibody complex, a biotin labeled TnI antibody complex and a fluorescently labeled quality control complex; and a reaction buffer solution containing a fluorescently labeled CK-MB antibody complex,a fluorescently labeled Myoglobin antibody complex and the fluorescently labeled quality control complex. The simultaneous detection of TnI, CK-MB and Myoglobin on the same reagent strip is achieved,the problems of poor precision and poor sensitivity during the single project test are solved, and the stability of the TnI / CK-MB / Myo kit is improved.

Description

technical field [0001] The invention relates to the technical field of fluorescent immunochromatography in medical immunology, in particular to a combined detection kit and detection method for cardiac troponin I, creatine kinase isoenzyme and myoglobin. Background technique [0002] At present, the common method for detecting cardiac troponin I / creatine kinase isoenzyme / myoglobin (TnI / CK-MB / Myo) in the market is fluorescent immunoassay, which is based on reagent strips with binding pads ( See figure 1 ). This reagent strip has a sample pad 1 , an absorbent pad 2 , a binding pad 3 and an NC membrane (nitrocellulose membrane) 4 . The NC membrane 4 has a quality control line 5 , a test line 6 , a test line 7 and a test line 8 . Because this method directly coats the fluorescently labeled TnI, CK-MB, Myoglobin antibody complex and quality control complex combined with the antigen on the binding pad of the reagent strip, when the sample flows through the binding pad, the The ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/72G01N33/68G01N33/573G01N33/558G01N33/58
CPCG01N33/72G01N33/6887G01N33/573G01N33/558G01N33/58G01N33/582G01N2333/4712G01N2333/805G01N2333/9123
Inventor 王洪涛李登红庄子槟梁健嘉李哉珉金永敏崔义烈
Owner 巴迪泰(广西)生物科技有限公司
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