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Carbon dioxide kit employing enzymatic cycling assay

A carbon dioxide and reagent kit technology, which is applied in biological testing, material inspection products, measuring devices, etc., can solve the problems of failure to effectively ensure the stability of reagent reagents, improvement of NADH stability of kits, improvement of kit stability, etc. , to achieve the effects of improving test stability, prolonging storage time, and low cost

Inactive Publication Date: 2018-04-27
ZHONGSHAN CHUANGYI BIOCHEM ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Patent CN2653492Y provides a carbon dioxide binding force tester; the test method used is the volumetric method, and the stability of the kit suitable for biochemical analyzers is not improved; patent CN200610097207.0 provides a preparation of a carbon dioxide kit method, but there is no relevant discussion on the recycling of NADH analogues, and the stability of the kit is poor in practical applications; CN103698500B provides a method for enhancing the stability of carbon dioxide detection reagents, but the main means are to add oily substances and surface active agent to reduce the contact between the reagent and carbon dioxide in the air, but the oily substances introduced may have other effects on the mechanical modules of the biochemical analyzer, and the NADH stability of the kit has not been improved; Discussion on the recycling of active substances, but does not involve the energy-supplying substances during recycling, and fails to effectively guarantee the stability requirements of reagents in use

Method used

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  • Carbon dioxide kit employing enzymatic cycling assay
  • Carbon dioxide kit employing enzymatic cycling assay
  • Carbon dioxide kit employing enzymatic cycling assay

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] The NADH analog regeneration system consists of the following components: 6-phosphate-glucose dehydrogenase 20U / L, hexokinase 200U / L, glucose 2g / L, adenosine triphosphate disodium salt 1g / L.

[0016] Fully dissolve and mix the above components into the original carbon dioxide measurement kit. The original carbon dioxide measurement kit consists of liquid single reagents: Trisbase 6g / L, hydrochloric acid 1.8g / L, stabilizer 1g / L, magnesium acetate 1g / L, sodium oxamate 0.3g / L, phosphoenolpyruvate Carboxylase 0.1g / L, Potassium Phosphoenolpyruvate 0.6g / L, APADH 0.5g / L, Malate Dehydrogenase 0.05g / L, Disodium EDTA 1g / L, Proclin300 1g / L L. Wherein, the stabilizer is at least one of gelatin, sorbitol, glycerol, ethylene glycol, and bovine serum albumin.

[0017] Using Landau Routine Biochemical Quality Control Level 2 (Quality Control 2) and Routine Biochemical Quality Control Level 3 (Quality Control 3) as samples, the standard product is Randox Routine Biochemical Calibratio...

Embodiment 2

[0022] The NADH analog cycle regeneration system consists of the following components: 6-phosphate-glucose dehydrogenase 5U / L, hexokinase 50U / L, glucose 1g / L, adenosine triphosphate disodium salt 0.5g / L .

[0023] Fully dissolve and mix the above components into the original carbon dioxide measurement kit. The original carbon dioxide measurement kit consists of liquid single reagents: Trisbase 6g / L, hydrochloric acid 1.8g / L, stabilizer, magnesium acetate 1g / L, sodium oxamate 0.3g / L, phosphoenolpyruvate carboxylase 0.1g / L, potassium phosphoenolpyruvate 0.6g / L, APADH 0.5g / L, malate dehydrogenase 0.05g / L, disodium edetate 1g / L, Proclin300 1g / L.

[0024] Detect according to the method of embodiment 1.

[0025] With contrast reagent stability performance index control result (table 2)

[0026]

Embodiment 3

[0028] The NADH analog cycle regeneration system consists of the following components: 6-phosphate-glucose dehydrogenase 50U / L, hexokinase 500U / L, glucose 5g / L, adenosine triphosphate disodium salt 2.5g / L .

[0029] Fully dissolve and mix the above components into the original carbon dioxide measurement kit. The original carbon dioxide measurement kit consists of liquid single reagents: Trisbase 6g / L, hydrochloric acid 1.8g / L, stabilizer, magnesium acetate 1g / L, sodium oxamate 0.3g / L, phosphoenolpyruvate carboxylase 0.1g / L, potassium phosphoenolpyruvate 0.6g / L, APADH 0.5g / L, malate dehydrogenase 0.05g / L, disodium edetate 1g / L, Proclin300 1g / L.

[0030] Detect according to the method of embodiment 1.

[0031] With contrast reagent stability performance index control result (table 3)

[0032]

[0033] From the data of the above three examples, it can be known that the carbon dioxide determination kit without the NADH analogue recycling system is compared with the kit with ...

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Abstract

The invention discloses a carbon dioxide kit employing enzymatic cycling assay. The kit is composed of a NADH analogue cyclic regeneration system and a carbon dioxide test reagent, wherein the NADH analogue cyclic regeneration system consists of 6-phosphogluconate dehydrogenase, hexokinase, glucose and adenosine triphosphate disodium. The carbon dioxide kit is simple to prepare and low in cost, and the NADH analogue cyclic regeneration system in the kit can prolong the storage time of the kit and effectively improves the stability, accuracy and precision of testing.

Description

technical field [0001] The invention relates to the field of in vitro diagnosis, in particular to a carbon dioxide kit using a cycle enzyme method. Background technique [0002] Total carbon dioxide (T-CO 2 ) refers to the various forms of CO in plasma 2 Total, including HCO 3 - (95%), a small amount of physically dissolved CO 2 and very small amounts of other forms of CO 2 . Therefore, it is affected by two factors of respiration and metabolism in the body, mainly the influence of metabolic factors. T-CO 2 Enzyme method, electrode method, titration method or volumetric method can be used. 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 substanc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50
CPCG01N33/5002
Inventor 崔海林黄小全李冰曾晓君黄清媚
Owner ZHONGSHAN CHUANGYI BIOCHEM ENG
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