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Method for simultaneous determination of double items of urea nitrogen and creatinine in serum

A determination method, urea nitrogen technology, applied in biochemical equipment and methods, determination/inspection of microorganisms, etc.

Inactive Publication Date: 2016-09-21
TIANJIN BAODI HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The determination of creatinine in clinical laboratory blood and urine is usually performed by enzymatic method or alkaline picric acid method (Jaffe method). Although the Jaffe method is relatively cheap, repeatable and accurate The sex is also good, but it is susceptible to the interference of other pseudo-creatinine substances in the serum, especially when the serum bilirubin value ≥ 165.5μmol / L, the negative deviation begins to appear, and the higher the serum bilirubin, the greater the deviation
In addition, Jaffe's method is severely interfered by ketone bodies, chylotemia, and hemolysis, and drugs such as cephalosporins, vitamin C, and dopamine also greatly interfere with the results

Method used

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  • Method for simultaneous determination of double items of urea nitrogen and creatinine in serum
  • Method for simultaneous determination of double items of urea nitrogen and creatinine in serum
  • Method for simultaneous determination of double items of urea nitrogen and creatinine in serum

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Experimental program
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Effect test

Embodiment 1

[0028] Composition of reagents:

[0029] a. Reagent I:

[0030] Reagent Ⅰ contains TRIS-hydrochloride phosphate buffer 50mmol / L, urease 5000U / L, glutamate dehydrogenase 9000U / L, α-ketoglutarate 160mmol / L, NADH 3.0mmol / L, Proclin-300 150-250 μl.

[0031] b. Reagent II:

[0032] Reagent II contains TRIS-hydrochloride phosphate buffer 50mmol / L, creatinine amidohydrolase 20KU / L, creatine amidinohydrolase 20KU / L, Proclin-300 150-250μl.

[0033] c. Standard solution: The standard solution is 7.14mmol / urea nitrogen.

Embodiment 2

[0035] Measurement procedure

[0036] Determination method: On the Japanese OLYMPUS AU2700 fully automated biochemical analyzer, the instrument automatically adds 3 μl of sample to 225 μl of reagent Ⅰ and mixes well, incubates at 37°C for 3 minutes, adds 75 μl of reagent Ⅱ, mixes well, and incubates at 37°C for 5.1 minutes, fully automatic analysis The instrument detects at a wavelength of 340nm. The instrument automatically calculates the results of urea nitrogen and creatinine, see Table 1 for details:

[0037] Table 1. Automatic biochemical analyzer test conditions of the present invention

[0038]

[0039] Concentration of urea nitrogen = F × ΔOD BUN / Δt

[0040] Creatinine concentration = F × ΔOD Cr / Δt

[0041] where ΔOD BUN / Δt is the rate of urea nitrogen production per unit time, ΔOD Cr / Δt is the rate of creatinine production per unit time, F is the correction factor, the measurement method is the rate method with standard, and its real-time response curve ...

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Abstract

The invention discloses a method for simultaneous determination of double items of urea nitrogen and creatinine in serum, and belongs to the method for testing a material through testing color changes of reaction results by using visible light; the technical scheme comprises that a reagent II only comprises effective components of creatinine amidohydrolase and creatine amidinohydrolase; a reagent I contains effective components of urease, glutamate dehydrogenase, alpha-ketoglutarate and NADH. The determination method comprises the steps: firstly, carrying out 37 DEG C warm bath of serum with the reagent I for 3-5 minutes; carrying out a reaction of urea in the serum with the reagent I to generate NAD+; adding the reagent II, carrying out 37 DEG C warm bath for 4-7 minutes, hydrolyzing creatinine with the creatinine amidohydrolase to generate creatine; making the creatine generate urea under the action of the creatine amidinohydrolase, and making the urea and the reagent I generate NAD+ under the action of urease; at the wavelength of 340 nm, comparing the reaction speed with that of a standard treated by the same way, determining the change of the first-step reaction NADH, namely the content of urea nitrogen in the serum, and determining the change of the second-step reaction NADH, namely the content of the creatinine in the serum.

Description

technical field [0001] The invention relates to a method for simultaneously measuring urea nitrogen creatinine content, which belongs to a method for measuring enzymes; or a method for testing materials by using visible light to produce color changes in the results of test reactions, in particular to a method for biochemical analysis. A fast and accurate method for the determination of blood urea nitrogen creatinine in serum. Background technique [0002] Urea is the main end product of human protein metabolism, and amino acid deamination produces NH 3 and C0 2 , the two synthesize urea in the liver, 0.3g of urea nitrogen is produced per gram of protein metabolism, and the nitrogen content in urea is 28 / 60. Usually, the kidney is the main organ for excreting urea. reabsorption, but the faster the urine flow rate in the renal tubule, the less the reabsorption, that is, the maximum clearance rate is reached. Like blood creatinine, in the early stage of renal damage, blood ur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/58C12Q1/34C12Q1/32
CPCC12Q1/58C12Q1/32C12Q1/34
Inventor 李立和丁弘魏志斌
Owner TIANJIN BAODI HOSPITAL
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