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Direct bilirubin assay method and kit

A determination method and technology for bilirubin, applied in the biological field, can solve the problems of high price of direct bilirubin oxidase, unfavorable large-scale application, expensive kits, etc., achieving excellent accuracy and repeatability, and safe use. High performance and low environmental pollution

Inactive Publication Date: 2019-03-29
武汉中太生物技术有限公司
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AI Technical Summary

Problems solved by technology

Each of these four methods has advantages and disadvantages. Among them, the oxidase method has good linearity and repeatability, and strong anti-interference ability, so it is an ideal detection method. However, due to the high price and high cost of direct bilirubin oxidase, The kit is expensive, which is not conducive to large-scale application

Method used

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  • Direct bilirubin assay method and kit
  • Direct bilirubin assay method and kit
  • Direct bilirubin assay method and kit

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Experimental program
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Embodiment

[0026] 1) Direct Bilirubin Assay Kit

[0027] Substrate reagent: 100mM citric acid buffer pH3.7, glucose 2mM, Proclin300 0.01%;

[0028] Enzyme reagent: 10mM phosphate buffer pH7.0, glucose oxidase 36KU / L, peroxidase 12KU / L, polyethylene glycol 6000 1%wt, Proclin300 0.0.01%wt.

[0029] 2) Kit linear detection

[0030] Prepare the direct bilirubin standard substance, and use the direct bilirubin assay kit in step 1) for detection. The detection method is to incubate the substrate reagent and the sample at 37°C for 5 minutes, read A1, add the enzyme reagent, and read after 5 minutes of reaction A2, calculate A2-A1 is the final reaction degree.

[0031] The reaction product is suitable for semi-automatic and automatic biochemical analyzer detection. Test parameters: wavelength 450nm, endpoint method, the ratio of reagent to sample volume is 24:1. Both substrate reagent and enzyme reagent were added in excess.

[0032] After using the kit of the present invention to draw a ca...

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Abstract

The invention discloses a direct bilirubin assay method and a kit. The direct bilirubin assay method utilizes an oxidase and a substrate thereof to react to form hydrogen peroxide, and then hydrogen peroxide and peroxidase react to release oxygen. Direct bilirubin in a solution is oxidized to biliverdin, and the absorption peak is decreased at 450 nm. The degree of decrease is recorded to calculate the direct bilirubin content. According to the invention, another oxidase which is cheap, easy to obtain and stable and can generate hydrogen peroxide and the substrate thereof are used to replace direct bilirubin oxidase; the cost is greatly reduced compared with the cost of an original direct bilirubin oxidase assay method; the experimental data acquired by the detection method proves that thesensitivity, accuracy and repeatability of the analysis are excellent; and the detection reagent has the advantages of high use safety, low environmental pollution and good practicability.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a direct bilirubin assay method and a kit. Background technique [0002] Direct bilirubin (DBIL), also known as conjugated bilirubin, is formed by combining indirect bilirubin with glucuronic acid after entering the liver by the action of intrahepatic glucuronosyltransferase. Direct bilirubin is soluble in water, reacts directly with azo reagents, and can be excreted through the kidneys with urine. The liver plays an important role in the metabolism of bilirubin, including the uptake, binding and excretion of indirect bilirubin in the blood by liver cells. The increase of serum direct bilirubin indicates that the excretion of bilirubin from the biliary tract is obstructed after being processed by liver cells. The determination of direct bilirubin is helpful for the diagnosis and differential diagnosis of jaundice types. Increased direct bilirubin: mainly seen in obstructive jaundi...

Claims

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Application Information

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IPC IPC(8): G01N33/72G01N21/31
CPCG01N21/31G01N33/728
Inventor 陈莹
Owner 武汉中太生物技术有限公司
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