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Creatinine content determination method and creatinine diagnosis kit

A technology for a diagnostic kit and a determination method, which is applied in the field of medical inspection and determination, can solve the problems of urgent, inconvenient testing, and reduced testing accuracy of reagents, and achieves the effects of easy operation, high specificity, and simple enzymatic method.

Inactive Publication Date: 2006-05-03
王尔中
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

One of its disadvantages is that while the system generates the reduced nicotinamide coenzyme required for the reaction, it also offsets some of the enzymes such as alanine aminotransferase, aspartate aminotransferase, urea, ammonia, creatinine, and carbon dioxide. activity, causing the accuracy of the reagent test to be greatly reduced
The second disadvantage is that the reagent must be reacted for a period of time before the formal test in order to produce enough reduced nicotinamide coenzyme, which results in slow results and brings great inconvenience to the test

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] Embodiment one (single agent)

[0080] The creatinine diagnostic kit of the present embodiment comprises:

[0081] Buffer 80mmol / l

[0082] Reduced coenzyme 0.2mmo / l

[0083] Phosphoenolpyruvate 2mmol / l

[0084] Creatinase 6000U / l

[0085] Creatinase 6000U / l

[0086] Urease 20000U / l

[0087] Phosphoenolpyruvate Carboxylase 10000U / l

[0088] Malate dehydrogenase 10000U / l

[0089] Stabilizer 50% (total volume)

[0090] 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 negative reaction.

[0091] After the sample and reagents are added and allowed to mix, the following reactions occur:

[0092] Creatinine + water creatininase creatine

[0093] Creatine + water creatinase Sarcosine + urea

[0094] Urea + water Urease bicarbonate + ammonium ion

[0095] b...

Embodiment 2

[0100] Embodiment two (two doses)

[0101] The creatinine diagnostic reagent of the present embodiment has:

[0102] Reagent I

[0103] Buffer 100mmol / l

[0104] Reduced coenzyme 0.25mmo / l

[0105] Phosphoenolpyruvate 5mmol / l

[0106] Creatinase 8000U / l

[0107] Urease 30000U / l

[0108] Phosphoenolpyruvate Carboxylase 15000U / l

[0109] Malate dehydrogenase 15000U / l

[0110] Stabilizer 50% (total volume)

[0111] Reagent II

[0112] Buffer 100mmol / l

[0113] Creatinase 8000U / l

[0114] Stabilizer 50% (total volume)

[0115] 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 negative reaction.

[0116] The specific measurement steps are:

[0117] Creatinine + water creatininase creatine

[0118] Creatine + water creatin...

Embodiment 3

[0125] Embodiment three (three doses)

[0126] The creatinine diagnostic reagent of the present embodiment is three doses, has:

[0127] Reagent I

[0128] Buffer 120mmol / l

[0129] Reduced coenzyme 0.3mmo / l

[0130] Phosphoenolpyruvate 8mmol / l

[0131] Stabilizer 20mmol / l

[0132] Reagent II

[0133] Buffer 120mmol / l

[0134] Creatinase 10000U / l

[0135] Urease 40000U / l

[0136] Phosphoenolpyruvate Carboxylase 20000U / l

[0137] Malate dehydrogenase 20000U / l

[0138] Stabilizer 50% (total volume)

[0139] Reagent III

[0140] Buffer 120mmol / l

[0141] Creatinase 10000U / l

[0142] Stabilizer 50% (total volume)

[0143] Set on the automatic biochemical analyzer: temperature 25°C, reaction time 20 minutes, test main wavelength 340nm, test sub-wavelength above 405nm, the volume ratio of the creatinine sample to be tested is 1 / 25, and the reaction direction is negative reaction.

[0144] The specific measurement steps are:

[0145] Creatinine + water creatininase C...

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Abstract

The invention relates to a method for measuring the creatinine content and a creatinine diagnosis agent box in the field of medical testing technology. The agent box comprises: buffer solution, reducing coenzyme, phosphoenolpyruvate, creatinine enzyme, atinase enzyme, urease, phosphoenolpyruvate carboxylase, malate dehydrogenase and stabilizer. It mixes the sample and the agent with a certain volume ratio to do enzyme coupling reaction; then it dispositions the end resting material on the chemical analyzer to detect the speed of the main wavelength light absorption to measure the content of the creatinine.

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...

Claims

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Application Information

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IPC IPC(8): G01N33/70G01N21/31G01N21/33G01N21/75G01N21/77G01N33/50
Inventor 王尔中
Owner 王尔中
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