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Detection method of bilirubin and detection kit

A detection method, bilirubin technology, applied in the preparation of test samples, color/spectral characteristic measurement, etc., can solve the problems of shortening the reaction time, interference, etc., and achieve a wide application range, easy operation, and simple instrument parameter setting Effect

Active Publication Date: 2015-04-15
潍坊泽成生物技术有限公司
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  • Abstract
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Problems solved by technology

[0005] The technical problem to be solved in the present invention is to provide a kind of detection method and detection kit of enzymatic rate method human serum bilirubin, for quantitative determination in vitro The concentration of bilirubin in human serum and plasma overcomes the defect that the existing detection method is susceptible to serum background interference. The detection method of the present invention uses an enzyme-catalyzed oxidation method and a rate measurement method to completely eliminate the interference of serum background , shorten the reaction time, easy to operate, accurate and reliable results, suitable for various biochemical analysis instruments

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  • Detection method of bilirubin and detection kit
  • Detection method of bilirubin and detection kit
  • Detection method of bilirubin and detection kit

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Embodiment 1

[0035] Embodiment 1, a kind of detection kit, comprises packing box, buffer solution, enzyme reagent, standard solution; Packing box is the outer packing of kit;

[0036] The buffer is 0.1mol / L Tris-HCl buffer containing 4mmol / L sodium cholate and 15mmol / L SDS, or 1.86g / L EDTA-Na 2 0.2mol / L phosphate buffer.

[0037] The enzyme reagent is bilirubin oxidase solution, the concentration is 1-20U / ml, the specific concentration is determined according to the volume ratio of the buffer solution, and the final enzyme concentration in the reaction solution is guaranteed to be 0.05-2U / ml.

[0038] The volume ratio of buffer solution to enzyme reagent is 20-3:1.

[0039] The standard solution is free bilirubin solution or taurine-conjugated bilirubin solution, with a concentration of 5-40 μmol / L, and is calibrated with the national standard.

[0040] The storage conditions and validity period are as follows: the original reagent is stored at 2-8°C, and the validity period is 12 months...

Embodiment 2

[0060] Embodiment 2, a kind of detection kit, except that the following is different from embodiment 1, all the other are identical with the detection kit in embodiment 1; The buffer solution is the Tris-HCl buffer solution of 0.1mol / L, contains 4mmol / L cholic acid Sodium and 15mmol / L SDS, the pH value is 8.0; the enzyme reagent is bilirubin oxidase solution, the concentration is 1U / ml, the volume ratio of buffer solution to enzyme reagent is 20:1; the standard solution is free bilirubin solution , with a concentration of 10 μmol / L, which was calibrated with the national standard assignment.

[0061] The above detection kit is used to detect total bilirubin, and the detection method is carried out according to the following steps: the absorbance decrease rate measurement step is carried out according to the single reagent mode operation, firstly, the buffer solution and the enzyme reagent are made into a reagent working solution, and it is stabilized for 10 minutes; then Accor...

Embodiment 3

[0079] Embodiment 3, a kind of detection kit, except that the following is different from embodiment 1, all the other are identical with the detection kit in embodiment 1; buffer solution is the Tris-HCl buffer solution of 0.1mol / L, contains 4mmol / L cholic acid Sodium and 15mmol / L SDS, the pH value is 8.2; the enzyme reagent is bilirubin oxidase solution, the concentration is 1U / ml, the volume ratio of buffer solution and enzyme reagent is 3:1; the standard solution is free bilirubin solution , with a concentration of 20 μmol / L, calibrated with national standard assignment.

[0080] The above detection kit is used to detect total bilirubin, and the detection method is carried out according to the following steps: the absorbance decrease rate measurement step is carried out according to the single reagent mode operation, firstly, the buffer solution and the enzyme reagent are made into a reagent working solution, and it is stabilized for 10 minutes; then According to the measur...

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Abstract

The invention discloses a detection method of bilirubin. A velocity measurement method is adopted; the reducing velocity of the absorbancy of a reaction system is directly proportional to the concentration of the bilirubin in a sample at the position with 440-465nm of wavelength; and the reducing velocity of the absorbancy within 30-180s is measured after delaying for 30-60s, so as to obtain the content of the bilirubin. The reaction velocity is measured by an enzyme velocity measurement method; reading is carried out during a linear phase in a reaction process; and interference of a background of a serum sample can be completely removed; the accuracy of a result is improved; the detection method is short in delay time, and short in reading duration; the entire reaction time is greatly shortened; the efficiency is improved; removal of the background of the serum sample does not need to be considered; the detection method is simple and convenient to operate, parameters of an instrument are simple to set, and a complicated computational formula is not needed; and either a dual-reagent model or a single-reagent mode can be adopted for measurement, and the detection method is suitable for various automatic and self-automatic biochemical analyzers and spectrophotometers, thus being wide in application range.

Description

technical field [0001] The present invention relates to a kind of detection method of bilirubin, specifically a kind of detection method and detection kit of human serum bilirubin by enzymatic rate method, which are used for in vitro quantitative determination of the concentration of bilirubin in human serum and plasma. The invention belongs to the field of medical clinical biochemical diagnostic reagents. Background technique [0002] Bilirubin is a complex linear organic compound composed of four pyrrole rings connected by a 2-methine bridge in the center of the molecule, and is a metabolite of heme. In blood, bilirubin exists in two forms: conjugated bilirubin and unconjugated bilirubin. The absorption peak of conjugated bilirubin is 422nm, and the absorption peak of unconjugated bilirubin is 459nm. Under normal physiological conditions, conjugated bilirubin forms a non-covalent reversible bond with albumin, but in the blood of some liver patients, bilirubin and albumin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/31G01N1/38
Inventor 孙希武秦英王永庆吐尔洪·买买提
Owner 潍坊泽成生物技术有限公司
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