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A low-temperature and high-efficiency carbon dioxide kit using a cyclic enzymatic method

A carbon dioxide, cyclic enzymatic method, used in biochemical equipment and methods, microbial determination/inspection, instruments, etc., can solve the problems of high cost, limited measurement range, poor stability, etc., and achieve low cost, high stability and high stability. Sensitive, easy to formulate effects

Active Publication Date: 2022-04-01
DALIAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

High cost, poor stability, and limited measurement range are the current CO 2 Major Issues with Diagnostic Kits

Method used

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  • A low-temperature and high-efficiency carbon dioxide kit using a cyclic enzymatic method
  • A low-temperature and high-efficiency carbon dioxide kit using a cyclic enzymatic method
  • A low-temperature and high-efficiency carbon dioxide kit using a cyclic enzymatic method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Low-temperature and high-efficiency use of carbon dioxide kits for cyclic enzymatic methods, as follows:

[0015] NADH analog cycle regeneration system is composed of 6-phosphate-fructose dehydrogenase 10U / L, hexokinase 100U / L, fructose 1g / L, adenosine triphosphate disodium salt 0.5g / L, the above groups Fully dissolve and mix in the original carbon dioxide measurement kit, which consists of liquid single reagent: Tris-Hcl 0.1mol / L (pH=8), stabilizer 1g / L, oxaloacetic acid 3g / L, phosphoenolpyruvate carboxylase 0.1g / L, potassium phosphoenolpyruvate 0.6g / L, low temperature malate dehydrogenase 0.05g / L, Triton-100 1g / L; among them, the kit The medium stabilizer is at least one of sorbitol, mannitol, and glycerol; wherein, 6-phosphate-fructose dehydrogenase is derived from marine low-temperature glycerol-producing Candida (Candidaglycerol), and hexokinase is derived from marine low-temperature short Lactobacillus brevis, phosphoenolpyruvate carboxylase from marine low-temp...

Embodiment 2

[0022] Low-temperature and high-efficiency use of carbon dioxide kits for cyclic enzymatic methods, as follows:

[0023] NADH analog cycle regeneration system is composed of 6-phosphate-fructose dehydrogenase 5U / L, hexokinase 50U / L, fructose 0.5g / L, adenosine triphosphate disodium salt 0.25g / L, the above The components are fully dissolved and mixed in the original carbon dioxide measurement kit. The original carbon dioxide measurement kit consists of liquid single reagents: Tris-Hcl 0.1mol / L (pH=8), stabilizer 1g / L, oxaloacetate 3g / L, phosphoenolpyruvate carboxylase 0.1g / L, potassium phosphoenolpyruvate 0.6g / L, malate dehydrogenase 0.05g / L, Triton-100 1g / L; among them, the stabilizer in the kit is sorbitol, mannitol, and glycerin at least one of .

[0024] Detected according to the method of Example 1, the results are shown in Table 2.

[0025] Table 2 and contrast reagent stability performance index control result

[0026]

[0027] As can be seen from Table 2, the NAD...

Embodiment 3

[0029] Low-temperature and high-efficiency use of carbon dioxide kits for cyclic enzymatic methods, as follows:

[0030]The NADH analog cycle regeneration system consists of 6-phosphate-fructose dehydrogenase 20U / L, hexokinase 200U / L, fructose 2g / L, adenosine triphosphate disodium salt 1g / L, and the above components are fully Dissolve and mix in the original carbon dioxide measurement kit, which consists of liquid single reagent: ris-Hcl 0.1mol / L (pH=8), stabilizer 1g / L, oxaloacetic acid 3g / L, Phosphoenolpyruvate carboxylase 0.1g / L, potassium phosphoenolpyruvate 0.6g / L, malate dehydrogenase 0.05g / L, Triton-100 1g / L; among them, the stabilizer in the kit It is at least one of sorbitol, mannitol and glycerin.

[0031] Detected according to the method of Example 1, the results are shown in Table 3.

[0032] Table 3 and contrast reagent stability performance index control result

[0033]

[0034] As can be seen from Table 3, the NADH analog cycle regeneration system in the c...

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Abstract

The invention relates to the field of in vitro diagnosis, in particular to a low-temperature and high-efficiency carbon dioxide kit using a cycle enzyme method. The kit is composed of NADH analog cyclic regeneration system and carbon dioxide determination reagent. The NADH analog cyclic regeneration system consists of 6-phosphate-fructose dehydrogenase 5-50U / L, hexokinase 50-500U / L, fructose 1- 5g / L, adenosine triphosphate disodium salt 0.5-2.5g / L, carbon dioxide determination reagent consists of Tris-hydrochloric acid 0.1mol / L (pH=8), stabilizer 1g / L, oxaloacetic acid 0.3g / L, composed of phosphoenolpyruvate carboxylase 0.1g / L, potassium phosphoenolpyruvate 0.6g / L, low temperature malate dehydrogenase 0.05g / L, Triton-100 1g / L. The carbon dioxide measuring kit of the present invention can be used at a lower temperature to detect the carbon dioxide concentration in a high-salt sample.

Description

technical field [0001] The invention relates to the field of in vitro diagnosis, in particular to a low-temperature and high-efficiency carbon dioxide kit using a cycle enzyme method. Background technique [0002] CO in blood 2 It is a common indicator for judging the acid-base metabolism in the human body. Its monitoring provides an important basis for the process and curative effect judgment of acid-base balance disorders, and is one of the indispensable emergency items. Serum total carbon dioxide is mostly measured clinically by titration and ion selective electrode (ISE). Among them, enzymatic determination of T-CO 2 After the reagent is opened and used, it is easy to absorb carbon dioxide in the air, causing the NADH substances in the reagent to be oxidized and invalidated. The electrode method is generally combined with an electrolyte analyzer. CO 2 The electrode membrane needs to be replaced periodically. In the enzymatic method, NADH substances are used as the re...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/527C12P19/36
CPCC12P19/36C12Q1/527G01N2333/988
Inventor 张庆芳肖景惠王梦雨迟乃玉
Owner DALIAN UNIV
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