Determination method of creatnine content and reagent box for diagnosing creatnine

Abstract

A method for measuring the content of creatinine by use of the creatinine testing reagent kit includes such steps as proportionally mixing said reagents with speciment, enzyme coupling reaction, taking the resultant, detesting the variation of primary wave in optical absorbancy by biochemical analyzer, and calculating the content of creatinine. Said reagent kit is composed 11 reagents including buffer liquid, adenosine triphosphate, oxidizing coenzyme, stabilizer, etc.

Description

technical field

[0001] The invention relates to a method for measuring creatinine content, and at the same time, the invention also relates to a creatinine diagnostic kit for realizing the method, which belongs to the technical field of medical examination and measurement. Background technique

[0002] Determination of creatinine mainly includes chemical determination method (Jaffe method), enzymatic method, high performance liquid chromatography and capillary electrophoresis.

[0003] Chemical determination method - low cost, easy to operate, is currently one of the most commonly used methods for the determination of creatinine in China. The creatinine in the specimen interacts with picrate to form a yellow-red complex of picric acid and creatinine. The disadvantage of this method is that the specificity is not high, because vitamin C, acetone, acetoacetic acid, methyldopa and high concentration of glucose, protein and some antibiotics such as penicillin G, cefoxitin, cefa...

Examples

Embodiment 1

[0088] Embodiment one (single dose)

[0089] The creatinine diagnostic kit of the present embodiment comprises:

[0090] Buffer 80mmol / l

[0091] Adenosine triphosphate 1mmol / l

[0092] Pyruvate 4mmol / l

[0093] Ethanol 2mmol / l

[0094] Oxidized coenzyme 1mmol / l

[0095] Creatinine Deiminase 2000U / l

[0096] N-Methylhydantoinase 2000U / l

[0097] Pyruvate oxidase 10000U / l

[0098] Catalase 10000U / l

[0099] Aldehyde dehydrogenase 10000U / l

[0100] Stabilizer 50% (total volume)

[0101] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, test main wavelength 340nm, test sub-wavelength above 405nm, the volume ratio of the creatinine sample to the reagent to be tested is 1 / 25, and the reaction direction is positive reaction.

[0102] After the sample and reagents are added and allowed to mix, the following reactions occur:

[0103] Creatinine + water + H + creatinine deiminase N-Methylhydantoin + ammonia ion

[0104] N-Meth...

Embodiment 2

[0112] Embodiment two (two doses)

[0113] The creatinine diagnostic reagent of the present embodiment has:

[0114] Reagent I

[0115] Buffer 100mmol / l

[0116] Adenosine triphosphate 3mmol / l

[0117] Pyruvate 7mmol / l

[0118] Ethanol 5mmol / l

[0119] Oxidized coenzyme 2mmol / l

[0120] N-Methylhydantoinase 5000U / l

[0121] Pyruvate oxidase 15000U / l

[0122] Catalase 15000U / l

[0123] Aldehyde dehydrogenase 15000U / l

[0124] Stabilizer 50% (total volume)

[0125] Reagent II

[0126] Buffer 100mmol / l

[0127] Creatinine deiminase 5000U / l

[0128] Stabilizer 50% (total volume)

[0129] When measuring creatinine content, the temperature is controlled at 30°C, the reaction time is 15 minutes, the main wavelength of the test is 340nm, the secondary wavelength of the test is above 405nm, the volume ratio of the creatinine sample to the reagent is 1 / 25, and the reaction direction is positive reaction.

[0130] The specific measurement steps are:

[0131] Creatinine...

Embodiment 3

[0140] Embodiment three (three doses)

[0141] The creatinine diagnostic reagent of the present embodiment is three doses, has:

[0142] Reagent I

[0143] Buffer 120mmol / l

[0144] Adenosine triphosphate 5mmol / l

[0145] Acetaldehyde + oxidized coenzyme + water Aldehyde dehydrogenase Acetate + reducing coenzyme + hydrogen ion

[0146] The final reactant is placed under a biochemical analysis instrument, and the increase of the absorbance at the main wavelength of 340nm is detected, so as to calculate the content of creatinine.

[0147] The reaction time of each reaction step was controlled at 20 minutes.

technical field

[0001] The invention relates to a method for measuring creatinine content, and at the same time, the invention also relates to a creatinine diagnostic kit for realizing the method, which belongs to the technical field of medical examination and measurement. Background technique

[0002] Determination of creatinine mainly includes chemical determination method (Jaffe method), enzymatic method, high performance liquid chromatography and capillary electrophoresis.

[0003] Chemical determination method - low cost, easy to operate, is currently one of the most commonly used methods for the determination of creatinine in China. The creatinine in the specimen interacts with picrate to form a yellow-red complex of picric acid and creatinine. The disadvantage of this method is that the specificity is not high, because vitamin C, acetone, acetoacetic acid, methyldopa and high concentration of glucose, protein and some antibiotics such as penicillin G, cefoxitin, cefa...

Examples

Embodiment 1

[0088] Embodiment one (single dose)

[0089] The creatinine diagnostic kit of the present embodiment comprises:

[0090] Buffer 80mmol / l

[0091] Adenosine triphosphate 1mmol / l

[0092] Pyruvate 4mmol / l

[0093] Ethanol 2mmol / l

[0094] Oxidized coenzyme 1mmol / l

[0095] Creatinine Deiminase 2000U / l

[0096] N-Methylhydantoinase 2000U / l

[0097] Pyruvate oxidase 10000U / l

[0098] Catalase 10000U / l

[0099] Aldehyde dehydrogenase 10000U / l

[0100] Stabilizer 50% (total volume)

[0101] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, test main wavelength 340nm, test sub-wavelength above 405nm, the volume ratio of the creatinine sample to the reagent to be tested is 1 / 25, and the reaction direction is positive reaction.

[0102] After the sample and reagents are added and allowed to mix, the following reactions occur:

[0103] Creatinine + water + H + creatinine deiminase N-Methylhydantoin + ammonia ion

[0104] N-Meth...

Embodiment 2

[0112] Embodiment two (two doses)

[0113] The creatinine diagnostic reagent of the present embodiment has:

[0114] Reagent I

[0115] Buffer 100mmol / l

[0116] Adenosine triphosphate 3mmol / l

[0117] Pyruvate 7mmol / l

[0118] Ethanol 5mmol / l

[0119] Oxidized coenzyme 2mmol / l

[0120] N-Methylhydantoinase 5000U / l

[0121] Pyruvate oxidase 15000U / l

[0122] Catalase 15000U / l

[0123] Aldehyde dehydrogenase 15000U / l

[0124] Stabilizer 50% (total volume)

[0125] Reagent II

[0126] Buffer 100mmol / l

[0127] Creatinine deiminase 5000U / l

[0128] Stabilizer 50% (total volume)

[0129] When measuring creatinine content, the temperature is controlled at 30°C, the reaction time is 15 minutes, the main wavelength of the test is 340nm, the secondary wavelength of the test is above 405nm, the volume ratio of the creatinine sample to the reagent is 1 / 25, and the reaction direction is positive reaction.

[0130] The specific measurement steps are:

[0131] Creatinine...

Embodiment 3

[0140] Embodiment three (three doses)

[0141] The creatinine diagnostic reagent of the present embodiment is three doses, has:

[0142] Reagent I

[0143] Buffer 120mmol / l

[0144] Adenosine triphosphate 5mmol / l

[0145] Acetaldehyde + oxidized coenzyme + water Aldehyde dehydrogenase Acetate + reducing coenzyme + hydrogen ion

[0146] The final reactant is placed under a biochemical analysis instrument, and the increase of the absorbance at the main wavelength of 340nm is detected, so as to calculate the content of creatinine.

[0147] The reaction time of each reaction step was controlled at 20 minutes.