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Reagent for creatine kinase activity test

A technology for creatine kinase and activity determination, which is applied in biological testing, microbial determination/inspection, color/spectral property measurement, etc., and can solve the problems of unsatisfactory CK activator effect and inability to perform accurate determination.

Active Publication Date: 2007-02-21
SYSMEX CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the effect of EDTA on stabilizing the CK activator is not very satisfactory, and the CK in the measurement sample cannot be sufficiently activated with a reagent that has been stored for a long time, and accurate measurement cannot be performed in some cases.

Method used

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  • Reagent for creatine kinase activity test

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The basic constitutions of the first reagent and the second reagent for measuring CK activity are listed below.

[0034] pH 6.6

[0035] Imidazole 125mM

[0036] Magnesium acetate 12.5mM

[0037] Cyclohexanediaminetetraacetic acid 2.0mM

[0038] ADP 2.5mM

[0039] AMP 6.25mM

[0040] AP5A 12.5M

[0041] Glucose 25mM

[0042] NADP 2.5mM

[0043] G6PDH 1875U / L

[0044] Glucokinase 3750U / L

[0045] NAC 30mM

[0046] pH 9.0

[0047] Phosphocreatine 150mM

[0048] The above substances were dissolved in distilled water at the above concentrations to prepare the first reagent and the second reagent A. Then add aminomethanesulfonic acid 5mM to the second reagent A to prepare the second reagent B; add aminomethanesulfonic acid 10mM to the second reagent A to prepare the second reagent C; add aminomethanesulfonic acid to the second reagent A 20mM, prepare the second reagent D; then prepare the second reagent E with the same composition as the second reagent A.

[0...

Embodiment 2

[0054] The basic configurations of the first reagent and the second reagent for measuring CK activity are as follows.

[0055] pH 6.6

[0056] Imidazole 125mM

[0057] Magnesium acetate 12.5mM

[0058] CyDTA 2.0mM

[0059] ADP 2.5mM

[0060] AMP 6.25mM

[0061] AP5A 12.5 μM

[0062] Glucose 30mM

[0063] NADP 2.5mM

[0064] G6PDH 1875U / L

[0065] Glucokinase 3750U / L

[0066] NAC 30mM

[0067] pH 9.0

[0068] Phosphocreatine 150mM

[0069] The above substances were dissolved in distilled water at the above concentrations to prepare the first reagent A and the second reagent. Then, add 1 mM aminomethanesulfonic acid to the first reagent A to prepare the first reagent B; then add 2 mM aminomethanesulfonic acid to the first reagent A to prepare the first reagent C.

[0070] Store the above-mentioned first reagents A to C and the second reagent at a temperature of 4°C for 1 month, then use Hitachi 7170S automatic analyzer to use 180 μl of the first reagent and 45 μl o...

Embodiment 3

[0075] The effect of aminomethanesulfonic acid on stabilizing SH compounds was evaluated using reagents for measuring CK activity containing SH compounds other than NAC.

[0076] The following substances were dissolved in distilled water at the following concentrations and the pH was maintained at 6.6.

[0077] Imidazole 125mM

[0078] Magnesium acetate 12.5mM

[0079] EDTA 2.5mM

[0080] ADP 2.5mM

[0081] AMP 6.25mM

[0082] AP5A 12.5 μM

[0083] Glucose 30mM

[0084] NADP 2.5mM

[0085] G6PDH 1875U / L

[0086] Glucokinase 3750U / L

[0087] One of 30mM cysteamine, dithiothreitol (DTT), cysteine, glutathione and thioglycerol and 10mM aminomethanesulfonic acid were added to the above ingredients as a CK activator to prepare 5 a first reagent. At the same time, one of 30 mM cysteamine, dithiothreitol (DTT), cysteine, glutathione, and thioglycerol was added as a CK activator, and aminomethanesulfonic acid was not added, and then prepared 5 first reagents. The SH groups...

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Abstract

This invention provides a CK (creatine kinase) activity-measuring reagent containing a substance capable of inhibiting the deterioration of a CK-activating agent in its reagent solution.

Description

technical field [0001] The present invention relates to a reagent for measuring the activity of creatine kinase (hereinafter referred to as CK) contained in a sample. Background technique [0002] CK is a dimeric phosphatase composed of two subgroups. There are two subgroups of CK, type B (brain type) and type M (muscle type). CK has three isoenzymes (CK-MM, CK-MB, and CK-BB) due to the different combinations of the two subgroups. CK-MM is mostly contained in the muscles and bones, CK-MB is mostly contained in the myocardium, and CK-BB is mostly contained in the myocardium. contained in the brain. Due to diseases such as myocardial infarction and muscular dystrophy, the CK isoenzyme present at the diseased site is lost into the blood. Therefore, the activity value of CK present in samples such as serum and plasma in clinical examinations is used to diagnose the above diseases. important indicators. [0003] CK will be inactivated in blood, and the activity measurement sh...

Claims

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Application Information

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IPC IPC(8): G01N33/00G01N21/31G01N33/53C12Q1/25
Inventor 酒井康裕
Owner SYSMEX CORP