Creatine kinase isozyme detection reagent kit and preparation method thereof
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A detection kit, creatine kinase technology, applied in measurement devices, instruments, scientific instruments, etc., can solve problems such as low sensitivity and too many interference factors, and achieve the effects of high sensitivity, wide application to a wide range of people, and strong practicability
Inactive Publication Date: 2019-04-12
深圳天辰医疗科技有限公司
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[0015] The main purpose of the present invention is to provide a creatine kinase isoenzyme detection kit, which aims to solve the technical problem of low sensitivity caused by too many interference factors in the existing CK-MB detection method
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[0049] On the other hand, the embodiment of the present invention also provides the preparation method of the creatine kinase isoenzyme detection kit described above, including:
[0050] a) React the magnetic particles with the first anti-CK-MB monoclonal antibody and the second anti-CK-MB monoclonal antibody to prepare the first anti-CK-MB monoclonal antibody and the second anti-CK-MB monoclonal antibody bound to the surface Antibody magnetic particles;
[0051] b) coupling the third anti-CK-MB monoclonal antibody or the fourth anti-CK-MB monoclonal antibody to a labeled enzyme to prepare an enzyme-labeled detection antibody;
[0052] Wherein, the binding activity of the second anti-CK-MB monoclonal antibody to the magnetic particles is better than that of the first anti-CK-MB monoclonal antibody to the magnetic particles;
[0053] The first anti-CK-MB monoclonal antibody and the second anti-CK-MB monoclonal antibody are covalently bound to the magnetic particles.
[0054] ...
Embodiment 1
[0095] This embodiment provides a creatine kinase isoenzyme detection kit, the preparation method of which comprises:
[0096] 1. Reagent preparation
[0097] Preparation of MES buffer: Take 21.32g of MES and add about 800mL of pure water to dissolve it, then adjust the pH to 5.0 with NaOH, and dilute to 1L.
[0098] Preparation of cross-linking reagent: Add appropriate amount of EDC to MES buffer, and adjust the concentration of EDC to 10 mg / mL.
[0099] Preparation of blocking solution: Weigh 6.05g of Tris and 9.0g of NaCl, add 80ml of pure water to dissolve them, then adjust the pH to 7.2 with HCl, add 5.0g of BSA, 1.0g of Tween-20, mix well, and set the volume to 1000mL.
[0100] Preparation of cleaning solution: Weigh 3.025g of Tris and 9g of NaCl, add about 800mL of pure water to dissolve them, adjust the pH to 7.2 with HCl, add 0.5g of Tween-20, mix well, and dilute to 1L.
[0101] Preparation of reagent R1: Take 6.5g of Tris and 9.0g of sodium chloride, add 80mL of p...
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Abstract
The invention belongs to the technical field of in-vitro detecting, and particularly relates to a creatine kinase isozyme detection reagent kit and a preparation method thereof. The creatine kinase isozyme detection reagent kit comprises a magnetic separation reagent and an enzyme labeled antibody; the magnetic separation reagent comprises magnetic particles, and a first anti-CK-MB monoclonal antibody and a second anti-CK-MB monoclonal antibody are combined with the surfaces of the magnetic particles; the enzyme labeled antibody comprises an enzyme labeled third anti-CK-MB monoclonal antibodyand/or an enzyme labeled fourth anti-CK-MB monoclonal antibody; and the first anti-CK-MB monoclonal antibody, the second anti-CK-MB monoclonal antibody, the third anti-CK-MB monoclonal antibody and the fourth anti-CK-MB monoclonal antibody are specific to different epitopes of creatine kinase isozyme. According to the creatine kinase isozyme detection reagent kit and the preparation method thereof, the surfaces of the magnetic particles in the magnetic separation reagent are combined with the first anti-CK-MB monoclonal antibody and the second anti-CK-MB monoclonal antibody which are specificto the different epitopes of the creatine kinase isozyme, multi-site recognition can be conducted on the CK-MB antigens in a to-be-detected sample, combination between a magnetic bead antibody and theantigens is improved, the detection missing situation can be avoided, the sensitivity is high, and the applicable people range is wide.
Description
technical field [0001] The invention belongs to the technical field of in vitro detection, and in particular relates to a creatine kinase isozyme detection kit and a preparation method thereof. Background technique [0002] Cardiovascular disease is a major fatal disease that endangers the health and life of the people in our country. According to the statistics of the Ministry of Health, in my country's cities, 235 people die of cardiovascular and cerebrovascular diseases for every 100,000 people, accounting for 100% of all diseases. The number of deaths caused by the disease is the first, and it is increasing at a rate of 2% every year. Acute myocardial infarction (AMI) refers to acute necrosis of part of the myocardium caused by persistent and severe myocardial ischemia, and is the main cause of death in cardiovascular patients. Clinically, it often manifests as chest pain, acute circulatory dysfunction, and a series of characteristic ECG changes reflecting myocardial inj...
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