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Reagent for determining creatine kinase and isoenzymes thereof and kit for determining creatine kinase and isoenzymes thereof

A creatine kinase and isoenzyme technology, applied in the field of biochemical detection, can solve the problems of inactivation, oxidation, and the accuracy of isoenzyme determination kits, etc., to prevent oxidative inactivation, prolong the service life, and ensure accurate sexual effect

Active Publication Date: 2019-03-26
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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  • Reagent for determining creatine kinase and isoenzymes thereof and kit for determining creatine kinase and isoenzymes thereof
  • Reagent for determining creatine kinase and isoenzymes thereof and kit for determining creatine kinase and isoenzymes thereof
  • Reagent for determining creatine kinase and isoenzymes thereof and kit for determining creatine kinase and isoenzymes thereof

Examples

Experimental program
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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 ...

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...

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Abstract

The invention provides a reagent for determining creatine kinase and isoenzymes thereof and a kit for determining the creatine kinase and isoenzymes thereof and belongs to the technical field of biochemical detection. The reagent for determining creatine kinase and isoenzymes thereof comprises ionic liquid 1-ethyl-3-methylimidazolium tetrafluoroborate. The kit for determining the creatine kinase (CK) and isoenzymes of the CK is composed of the ionic liquid 1-ethyl-3-methylimidazolium tetrafluoroborate, an activating agent, a chelating agent, a preservative, a buffer solution and the reagent for determining the CK and isoenzymes of the CK. The reagent and kit disclosed by the invention can be applied to a fully automatic biochemical analyzer, the operation is simple and convenient, the determination accuracy of the reagent can also be ensured after the reagent for determining the CK and isoenzymes of the CK is preserved for a long time, and the service life of the reagent can be obviously prolonged.

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 Applications(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