Quantitative creatine kinase-myoglobin (CK-MB) determination kit and assay method thereof

A technique for quantitative determination of creatine kinase is applied in the field of kits for determining the content of creatine kinase isoenzyme in human serum, which can solve the problems of short recovery time and early peak time of CK-MB.

Inactive Publication Date: 2012-05-02
INNER MONGOLIA KEHUI BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(3) There is some controversy in evaluating the infarct size of AMI by the serum CK-MB level. It is generally believed that the smaller the infarct size, the earlier the CK-MB peak time and the shorter the recovery time
3. CK-MB is not completely specific to myocardium, but also exists in a small amount in skeletal muscle

Method used

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  • Quantitative creatine kinase-myoglobin (CK-MB) determination kit and assay method thereof
  • Quantitative creatine kinase-myoglobin (CK-MB) determination kit and assay method thereof
  • Quantitative creatine kinase-myoglobin (CK-MB) determination kit and assay method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] 1. Operating procedures for preparing magnetic bead buffer: see Table 1 for the formula, taking the preparation of 1L as an example:

[0079] 1. Weigh 4.58g of TRIS and 6.81g of NaCl in a 1L container, weigh 0.96g of TWEEN-20 in a 20ml container, add an appropriate amount of water to dissolve it completely, and then pour it into the above container;

[0080] 2. Use a pipette to take 0.2ml of Proclin-300 and dissolve it completely in a beaker of a small amount of purified water, then pour it into the above-mentioned 1L container; then add 800ml of purified water into the 1L container, stir well to completely dissolve the reagent;

[0081] 3. Adjust the PH meter to measure the PH value; use HCl or NaOH to adjust the PH value, and measure the PH value between 7.95-8.05 to meet the requirements;

[0082] 4. Weigh 3g of BSA (bovine serum albumin) and pour it into the above container;

[0083] 5. Finally, set the volume to 1000ml, measure the pH value, the range is between 7...

Embodiment 2

[0100] 1. Operating procedures for the preparation of enzyme reactant diluent: the formula is shown in Table 2, taking the preparation of 1L as an example:

[0101] 1. Take Tris 6.06g, NaCl 13.0g, Zncl 2 0.05g, Proclin-300 0.2ml and MgCl 2 0.05g in the flask; then add 800ml of purified water into the flask and stir thoroughly to dissolve the reagent completely;

[0102] 2. Use HCl or NaOH to adjust the pH, and measure the pH to be in the range of 7.35-7.45;

[0103] 3. Weigh 3g of BSA and pour it into the above container;

[0104] 4. Finally, adjust the volume to 1000ml, measure the pH value, and if the range is between 7.35-7.45, it meets the requirements, and filter with a 0.2um filter; after filtering, label it and store it in a cold storage at 2-8°C, with a validity period of 12 months;

[0105] Enzyme reactant diluent Table 2

[0106]

[0107] 2. Coupling of alkaline phosphatase (ALP) and anti-CK-MB monoclonal antibody

[0108] 1. Add 10 mg of ALP to 5 ml of norma...

Embodiment 3

[0115] Reaction enhancer preparation operating procedures: see the formula (Table 3), taking the preparation of 1L as an example:

[0116] 1. Weigh 1.56g of TRIS (trihydroxymethylaminomethane, molecular formula: (HOCH2)3CNH2) and 4.23g of NaCl in a 1L container; use a pipette to measure 0.2ml of Proclin-300 into a beaker of a small amount of purified water After completely dissolved, pour into the above 1L container;

[0117] 2. Use a graduated cylinder to measure 800ml of purified water into a 1L container, stir well until it is completely dissolved; adjust the pH with HCL or NaOH, and measure the range between 7.35-7.45;

[0118] 3. Weigh 0.9g of Mak33 into the above 1L container;

[0119] 4. Finally, set the volume to 1000ml. After completely dissolving, measure the PH value. If the range is between 7.35-7.45, it meets the requirements. Filter with a 0.2um filter; after filtering, label it and store it in a cold storage at 2-8°C. month;

[0120] Preparation of reaction e...

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Abstract

The invention discloses a quantitative creatine kinase-myoglobin (CK-MB) determination kit. The kit is characterized in that the kit contains CK-MB magnetic separation reagent, enzymatic reactant, reaction enhancer, diluent, CK-MB calibrator, CK-MB quality control material, cleaner concentrate and substrate solution. The invention also discloses a preparation method for the kit. The kit integrates the chemiluminescence technology with immunomagnetic beads to provide a homogeneous phase-approximating reaction system. Compared with the prior art, the kit has higher assay sensitivity and specificity, and achieves better performance parameters, and mover, the product cost is greatly reduced.

Description

technical field [0001] The invention relates to a kit for measuring serum and a testing method thereof, in particular to a kit for measuring the content of creatine kinase isoenzyme (CK-MB) in human serum and a testing method thereof. Background technique [0002] The molecular weight of human creatine kinase isoenzyme is 85745. CK-MB is mainly distributed in the myocardium. When the myocardium is injured, CK-MB is released into the blood, and starts to increase after 3-8 hours, and can last for 3-5 days. The combined detection of troponin, myoglobin, and creatine kinase isoenzyme can detect myocardial damage this morning. CK is a dimer composed of M and B subunits, and two different subunits make up three isozymes. CK-BB (CK1) mainly exists in brain tissue CK-MB (CK2) mainly exists in cardiac muscle CK-MM (CK3) mainly exists in skeletal muscle tissue In addition, there is an isoenzyme (CK-MiMi) in mitochondria. The content of CK-MB in different parts of the myocardium i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N21/76
Inventor 不公告发明人
Owner INNER MONGOLIA KEHUI BIOLOGICAL TECH
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