Creatinine content detecting reagent and kit, and manufacturing and using methods of kit

A detection reagent, creatinine technology, applied in the field of creatinine detection, can solve the problems of overestimation of creatinine concentration, complex operation, poor selectivity, etc., and achieve high anti-interference and high sensitivity effects

Active Publication Date: 2012-03-14
TIANJIN HEOWNS BIOCHEM TECH
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Problems solved by technology

Mass spectrometry and liquid chromatography cannot be automated, the operation is complicated, and requires the analysis team to operate and maintain the instrument, while the ELISA method requires the use

Method used

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  • Creatinine content detecting reagent and kit, and manufacturing and using methods of kit
  • Creatinine content detecting reagent and kit, and manufacturing and using methods of kit
  • Creatinine content detecting reagent and kit, and manufacturing and using methods of kit

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Embodiment Construction

[0038] Below in conjunction with the examples, the present invention is further described, the following examples are illustrative, not limiting, and the protection scope of the present invention cannot be limited by the following examples.

[0039] The percentages mentioned in the following examples are percentages by weight unless otherwise specified, and the unit mM represents millimoles per liter.

[0040] 1. The chemical principle of creatinine kit

[0041]The creatinine kit described in the present invention includes four enzymes and one fluorescent dye, which are respectively: creatinase, creatinase, sarcosine oxidase and horseradish peroxidase. The chemical name of the fluorescent dye is 5(6)-carboxy-2,7-dichlorodihydrofluorescein dipivalate (structure as follows A), the fluorescent yield of the fluorescent dye is very low, so under general conditions, Its fluorescence emission efficiency is very low, and this fluorescent dye can be oxidized into 2,7-dichlorofluoresce...

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Abstract

The invention relates to a creatinine content detecting reagent and kit and manufacturing and using methods of the kit. The creatinine content detecting reagent comprises creatininase, creatinase, creatine oxidase and horseradish peroxidase; and a fluorescent dye, i.e., 5(6)-carboxyl-2,7-dichlordihydro fluorescein ester dipivalate comprises a dye bottom layer, a protease fixing layer and a top layer protecting layer. The using method has high selectivity; a protease has high specificity to reactants; the creatininase can be only used for catalyzing the hydrolysis reaction of creatinine, and does not have catalysis activity on other reaction substrates; three enzymes with high specificity are related to the detection process, so that the selectivity of the method is increased exponentially; the response fed by the kit is fluorescent response; and compared with certain methods for detecting color variations, the invention has the advantages of high sensitivity and interference resistance under the action of the inherent characteristics of fluorescent response.

Description

technical field [0001] The invention belongs to the field of creatinine detection, in particular to a creatinine content detection reagent, a kit, and a method for making and using the kit. Background technique [0002] Creatinine is a product of creatine phosphate metabolism, and its chemical structure is a lactam formed by dehydration condensation of creatine. Creatinine is generally present in the blood, passes through the blood circulation to the kidneys and is filtered out. For people with normal kidney function, the concentration of creatinine in the blood varies from person to person. Generally speaking, in the case of no heavy physical burden, the blood creatinine concentration of women is 45-60 μM, and the blood creatinine concentration of men fluctuates within the range of 60-110 μM. When there is a problem with renal function, the ability of the kidney to filter and remove creatinine will decrease, resulting in an abnormal increase in the concentration of creati...

Claims

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Application Information

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IPC IPC(8): G01N33/70G01N21/64
Inventor 徐维琛朱进清高玉兰程志敬
Owner TIANJIN HEOWNS BIOCHEM TECH
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