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Creatine kinase and its isozyme assay reagent and kit

A kit and reagent technology, applied in the field of biochemical detection, can solve the problems of unsatisfactory EDTA effect, poor accuracy of isoenzyme assay kit, loss of activity, etc.

Active Publication Date: 2021-09-10
WHITMAN BIOTECH NANJING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the mercapto group is extremely active in the reagent, and it is easily oxidized by oxygen or metal ions during storage and loses its activity. Usually, the chelating agent EDTA is used to chelate the metal ions that are easy to deteriorate, but the effect of EDTA is not ideal. In long-term storage , the activator still cannot fully activate CK, which is the main reason for the poor accuracy of CK and its isozyme assay kits during long-term storage, and is the most important factor affecting the application of reagents in the market

Method used

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  • Creatine kinase and its isozyme assay reagent and kit
  • Creatine kinase and its isozyme assay reagent and kit

Examples

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Embodiment 1

[0051] A kit for measuring CK, comprising a first reagent and a second reagent, the first reagent containing ionic liquid 1-ethyl-3-methylimidazolium tetrafluoroborate, imidazole buffer, NAC, EDTA , sodium azide, AP5A (diadenosine pentaphosphate), AMP (adenosine monophosphate), magnesium acetate, glucose, NADP+ (nicotinamide adenine dinucleotide phosphate) and glucose-6-phosphate dehydrogenase; The second reagent contains imidazole buffer, ADP (adenosine diphosphate), creatine phosphate, sodium azide and glucokinase.

[0052] The specific raw materials and proportions of this kit are as follows:

[0053] The composition and concentration of the first reagent of table 1

[0054] imidazole buffer 100mmol / L NAC 20mmol / L 1-Ethyl-3-methylimidazolium tetrafluoroborate 0.001％ EDTA 2mmol / L AP5A 12umol / L AMP 5mmol / L Glucose-6-phosphate dehydrogenase 3KU / L Magnesium acetate 10mmol / L glucose 25mmol / L NADP+ 3mmol / L...

Embodiment 2

[0065] A test kit for CK-MB determination, comprising a first reagent and a second reagent, the first reagent contains ionic liquid 1-ethyl-3-methylimidazolium tetrafluoroborate, imidazole buffer, NAC , EDTA, magnesium acetate, CK-M antibody, ADP, AMP, AP5A, glucose, glucose-6-phosphate dehydrogenase and sodium azide; the second reagent contains imidazole buffer, creatine phosphate, glucokinase, NADP+ and sodium azide.

[0066] The specific raw materials and proportions of this kit are as follows:

[0067] The composition and concentration of the first reagent of table 3

[0068]

[0069]

[0070] The composition and concentration of the second reagent of table 4

[0071] imidazole buffer 100mmol / L Glucokinase 3mmol / L creatine phosphate 165mmol / L NADP+ 2mmol / L Sodium azide 10mmol / L

[0072] In this embodiment, CK-MB in the sample is detected, and the specific steps are as follows:

[0073] (1) Mix 200uL of the first reagent...

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Abstract

The invention provides a creatine kinase and its isozyme assay reagent and its kit, which belong to the technical field of biochemical detection. The creatine kinase and its isozyme assay reagent contains ionic liquid 1-ethyl-3-methylimidazole tetra Fluoroborate, the kit for CK or CK isoenzyme determination consists of ionic liquid 1-ethyl-3-methylimidazolium tetrafluoroborate, activator, chelating agent, preservative, buffer and CK or CK isoenzyme activity assay reagent composition. The invention can be used for automatic biochemical analyzer detection, is easy to operate, can ensure the accuracy of reagent determination after long-term storage of CK and its isoenzyme determination reagents, and can significantly prolong the service life of the reagents.

Description

technical field [0001] The invention belongs to the technical field of biochemical detection, and in particular relates to a creatine kinase and its isozyme assay reagent and kit. Background technique [0002] Creatine Kinase (Creatine Kinase, CK) is a key enzyme in cell energy metabolism, and its activity measurement is the diagnostic basis for clinical skeletal muscle diseases and myocardial diseases. CK is composed of M and B subunits, and there are three subunits (isoenzyme) combinations CK-MM, CK-MB, and CK-BB, which are mainly distributed in skeletal muscle, cardiac muscle, and brain tissue of the human body. CK and CK-MB are important markers of conventional myocardial infarction. The normal reference value of CK in serum is 18.0-198.0 U / L, and the normal reference value of CK-MB is <0.6ng / ml. [0003] CK and its isoenzyme will be inactivated in blood, and its activity is unstable. It can only be stored for 4 hours at room temperature, and can only be stored for 8...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/48C12Q1/32
CPCC12Q1/32C12Q1/485G01N2333/904G01N2333/91215G01N2333/9123
Inventor 孙文勇郑佳费云燕常俊骏梁芬
Owner WHITMAN BIOTECH NANJING
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