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Creatine jubase MB isozyme activity detection reagent and preparation method thereof

A technique for measuring creatine kinase and activity, which is applied in the field of medical testing and can solve problems such as unreliability of measurement results

Active Publication Date: 2011-08-17
浙江东瓯诊断产品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The patent application number 200610114997.9 measures the activity of creatine kinase MB isoenzyme by reaction to generate hydrogen peroxide + luminol + peroxidase chemiluminescence method. It is easy to inactivate, and sulfhydryl reagents are usually added to maintain its activity. Sulfhydryl reagents will seriously inhibit the reaction of hydrogen peroxide + peroxidase, making the measurement results unreliable

Method used

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  • Creatine jubase MB isozyme activity detection reagent and preparation method thereof
  • Creatine jubase MB isozyme activity detection reagent and preparation method thereof
  • Creatine jubase MB isozyme activity detection reagent and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] The creatine kinase MB isoenzyme activity assay reagent of the present embodiment has the following main components and concentrations:

[0079] Imidazole buffer pH 6.7 100 mmol / L

[0080] Phosphocreatine 30 mmol / L

[0081] ADP 2 mmol / L

[0082] Glucose 20 mmol / L

[0083] GluK 2 KU / L

[0084] G6PDH 1.5 KU / L

[0085] 6PGDH 10 KU / L

[0086] Thiooxidized nicotinamide adenine dinucleotide phosphate 2 mmol / L

[0087] NAC 20 mmol / L

[0088] Anti-CK-M antibody (CK-M inhibition >2000 U / L)

[0089] EDTA 2mmol / L

[0090] Magnesium acetate 10 mmol / L

[0091] AMP 5 mmol / L

[0092] P 1 ,P 5 -DiAP 0.01 mmol / L

[0093] The creatine kinase MB isoenzyme activity assay reagent prepared in the present embodiment was measured on a Hitachi 7180 biochemical analyzer for precision test: 12 μL of the specimen to be tested was added to 300 μL of the creatine kinase MB isozyme activity assay reagent of the present invention In the process, mix well, incubate at 37°C for 1-3 minutes...

Embodiment 2

[0111] The creatine kinase MB isoenzyme activity assay reagent of the present embodiment has the following main components and concentrations:

[0112] Imidazole buffer pH 6.7 100 mmol / L

[0113] Phosphocreatine 30 mmol / L

[0114] ADP 2 mmol / L

[0115] Glucose 20 mmol / L

[0116] GluK 2 KU / L

[0117] G6PDH 2 KU / L

[0118] 6PGDH 15 KU / L

[0119] Thiooxidized nicotinamide adenine dinucleotide 2 mmol / L

[0120] NAC 20 mmol / L

[0121] Anti-CK-M antibody (CK-M inhibition >2000 U / L)

[0122] EDTA 2mmol / L

[0123] Magnesium acetate 10 mmol / L

[0124] AMP 5 mmol / L

[0125] P 1 ,P 5 -DiAP 0.01 mmol / L

[0126] The creatine kinase MB isoenzyme activity assay reagent prepared by the present embodiment is measured on a Hitachi 7180 biochemical analyzer for precision test: 12 μ L of the specimen to be tested is added to the creatine kinase MB isozyme activity assay reagent of the present invention In 300 μL, mix well, incubate at 37°C for 1 to 3 minutes, continuously monitor the ...

Embodiment 3

[0141] The creatine kinase MB isoenzyme activity assay reagent of the present embodiment is a double reagent, and its main components and concentrations are as follows:

[0142] Reagent 1:

[0143] Imidazole buffer pH 6.5 100 mmol / L

[0144] ADP 2 mmol / L

[0145] Glucose 20 mmol / L

[0146] GluK 2 KU / L

[0147] G6PDH 1.5 KU / L

[0148] 6PGDH 10 KU / L

[0149] Thiooxidized nicotinamide adenine dinucleotide phosphate 2 mmol / L

[0150] Thioglycerol 100 mmol / L

[0151] Anti-CK-M antibody (CK-M inhibition >2000 U / L)

[0152] EDTA 2mmol / L

[0153] Magnesium acetate 10 mmol / L

[0154] AMP 5 mmol / L

[0155] P 1 ,P 5 -DiAP 0.01 mmol / L

[0156] Reagent 2:

[0157] Tris / Hcl buffer pH 9.0 25 mmol / L

[0158] Phosphocreatine 150 mmol / L

[0159] The creatine kinase MB isoenzyme activity assay dual reagent prepared in this example was measured on a Hitachi 7180 biochemical analyzer for precision test: 8 μL of the specimen to be tested was added to 160 μL of reagent 1, mixed evenl...

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Abstract

The invention discloses a creatine jubase MB isozyme activity detection reagent which comprises sulfide-oxidizing coenzyme, 6-phosphaogluconate dehydrogenase, sulfhydryl reagent, magnesium salt, glucokinase or hexoxinase, anti-CK-M antibody, phosphocreatine, adenosine diphosphate, glucose and glucose-6-phosphate dehydrogenase. The preparation method of the reagent is as follows: dissolving all components in a buffer solution with pH of 5.0 to 9.5. The creatine jubase MB isozyme activity detection reagent can enlarge detection sensitivity several times; and the determination result has high precision and degree of accuracy.

Description

technical field [0001] The invention relates to an analytical reagent in the field of medical testing, in particular to a reagent for measuring the activity of creatine kinase MB isoenzyme in blood. Background technique [0002] Creatine kinase (CK) has three major isozymes, namely CK-MM, CK-MB, and CK-BB. Creatine kinase MB isoenzyme (CK-MB), which mainly exists in the myocardium, is the most specific enzyme in the myocardial enzyme spectrum. Since the 1980s, it has been recognized as the "gold standard" for myocardial enzyme diagnosis. Early diagnosis of acute myocardial infarction (AMI) is a sensitive and specific indicator for diagnosis and monitoring of AMI patients. [0003] Immunosuppression is the most commonly used clinically. The principle is: the anti-CK-M antibody in the immunosuppression reagent inhibits the CK-M subunit, that is, inhibits the activity of CK-MM and the activity of the CK-M subunit of CK-MB. Measure the activity of the uninhibited CK-B monomer ...

Claims

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Application Information

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IPC IPC(8): C12Q1/50
Inventor 不公告发明人
Owner 浙江东瓯诊断产品有限公司
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