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Double reagent method for detecting indirect bilirubin kit and preparation method

A technology of connecting bilirubin and double reagents, which is applied in the field of kits for measuring bilirubin in serum, plasma or urine, can solve the problems of error, workload and reagent cost doubling, red blood cell destruction, etc., to increase reagent cost , economical, convenient, and highly accurate

Inactive Publication Date: 2013-02-27
李立和
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  • Description
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AI Technical Summary

Problems solved by technology

3. Blood transfusion of incompatible blood type When incompatible blood is transfused, it will cause hemolysis and destroy a large number of red blood cells in the body, resulting in high indirect bilirubin in the blood
[0009] The direct bilirubin is determined by not adding dimethyl sulfoxide to the total bilirubin, and the determination is the direct bilirubin. At present, the methods for determining the indirect bilirubin in the laboratory are to measure the total bilirubin first, and then in the Determination of direct bilirubin, the subtraction of the two results is the value of indirect bilirubin, but this method doubles the workload and reagent cost, and is affected by two experimental errors at the same time, therefore, the present invention creates a double-reagent method A method for measuring indirect bilirubin at one time

Method used

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  • Double reagent method for detecting indirect bilirubin kit and preparation method
  • Double reagent method for detecting indirect bilirubin kit and preparation method
  • Double reagent method for detecting indirect bilirubin kit and preparation method

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Experimental program
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Effect test

Embodiment 1

[0026] Preparation of reagents:

[0027] a. Reagent A: Weigh 10.96 g of p-aminobenzenesulfonic acid and dissolve it in 800 ml of distilled water, add 18.83 ml of 36% concentrated hydrochloric acid, add 200 μL of Proclin-300, make up 1 liter of p-aminobenzenesulfonic acid 63.0 mmol / L, hydrochloric acid 294.0mmol / L, 0.2% Proclin-300.

[0028] b. Reagent B: Weigh 86.94 g of sodium nitrite and dissolve it in 1 liter of distilled water, add 200 μL of Proclin-300 to obtain 1260.0 mmol / L of sodium nitrite, 0.2% Proclin-300.

[0029] c. Reagent II: Weigh 1.87 g of dimethyl sulfoxide and dissolve it in 1 liter of distilled water, add 200 μL of Proclin-300 to obtain 24.0 mmol / L of dimethyl sulfoxide (DMSO).

[0030] Method of use: first mix reagent A and reagent B at 20:1 to form reagent I, mix reagent I with the sample and incubate for 3 minutes, then add reagent II for 5.1 minutes, measure the wavelength at 540nm, and perform colorimetry by the end point method, with the first The ...

Embodiment 2

[0033] In reagent II, dimethyl sulfoxide was changed to caffeine and sodium benzoate or glucuronosyltransferase. The content remains unchanged, and other ingredients remain unchanged.

Embodiment 3

[0035] Measurement procedure

[0036] Two-reagent method: On the Japanese OLYMPUS AU 2700 fully automated biochemical analyzer, the instrument automatically adds 5 μl of sample to 150 μl of reagent I and mixes, incubates at 37°C for 3 minutes, adds 50 μl of reagent II and mixes, and incubates at 37°C for 5.1 minutes. The automatic analyzer detects at a wavelength of 540nm, and the instrument automatically calculates the result of indirect bilirubin. See Table 1 for details

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

[0038]

[0039] OD IBIL =OD 27 -OD 10 ×[(SV+R I V I ) / (SV+R I V I +R II V II )]

[0040] Indirect bilirubin concentration = F × OD IBIL

[0041] where OD IBIL is the absorbance produced by indirect bilirubin, OD 10 is the absorbance measured after the sample is added to the working solution for reaction, OD 27 is the absorbance measured after adding reagent II to the sample, SV is the volume of the ...

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Abstract

The invention discloses a double reagent method for detecting indirect bilirubin in serum, which belongs to a material detecting method based on color change in the result of a test reaction by means of visible light. The technical scheme is that a diazo reagent is added in a reagent I, and the reagent I only contains active ingredients of an accelerator. The detecting method comprises the following steps: before use, a reagent A is mixed with a reagent B in the volume ratio of 20:1 to obtain the reagent I, warm bath is performed firstly on serum and the reagent I at 37 degrees C for 3 to 5 minutes, and direct bilirubin in the serum reacts with diazo acid chloride in the reagent I to obtain azobilirubin; then, a reagent II is added, warm bath is performed at 37 degrees C for 4 to 7 minutes; under the action of the accelerator, hydrogen bonds of the indirect bilirubin are broken, and then, the indirect bilirubin reacts with the diazo acid chloride to obtain the azobilirubin. The instrument is detected at the wavelength of 540 to 550 nm, and the azobilirubin obtained by the reaction of the reagent I is blank. Therefore, the content of the indirect bilirubin is calculated based on the azobilirubin obtained by the reaction of the reagent II.

Description

technical field [0001] The invention relates to a kit for measuring bilirubin in serum, plasma or urine; 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 detecting serum with a biochemical analyzer A two-reagent method for the determination of intermediate indirect bilirubin. Background technique [0002] Indirect bilirubin (abbreviated as IBIL) is mainly derived from the destruction of red blood cells, and the one that has not undergone glucuronidation in the liver is called indirect bilirubin. Indirect bilirubin can be converted into direct bilirubin after hepatic metabolism, which is discharged into the biliary tract with bile, and finally excreted through the stool. [0003] Clinical significance: Indirect bilirubin reference value: 1.0-20.0 μmol / L. Reasons for high indirect bilirubin: 1. Liver diseases Some malignant diseases can also cause high indirect bilirubin in the b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/72G01N21/31
Inventor 李立和
Owner 李立和
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