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Method for detecting hoptoglobin in serum and detecting kit thereof

A technology of haptoglobin and reagents, applied in the field of immunological detection, can solve the problems of slow display results, failure to widely carry out, large errors, etc., and achieve the effect of simple and easy detection method

Inactive Publication Date: 2011-07-20
BIOSINO BIO TECH & SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The operation steps of this method are complex and cumbersome, and the display result is slow and the error is large
The result shown is the content of hemoglobin, which only indirectly reflects the content of haptoglobin, and it has not been widely used clinically.

Method used

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  • Method for detecting hoptoglobin in serum and detecting kit thereof
  • Method for detecting hoptoglobin in serum and detecting kit thereof
  • Method for detecting hoptoglobin in serum and detecting kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 1. Preparation of reagents: including reagent R1 and reagent R2.

[0036] Reagent R1 contains: Tris 0.05M, polyethylene glycol 6000 2.5w / v%, Tween 20 0.2w / v%, sodium chloride 0.85w / v%, and the pH value is 8.0.

[0037] Reagent R2 contains: 0.05M trishydroxymethylaminomethane, 0.85w / v% sodium chloride, 35w / v% haptoglobin antibody, and the pH value is 8.0.

[0038] 2. Drawing of standard curve:

[0039] Use 6 concentrations of calibrator, the calibration concentration is X 1 、X 2 、X 3 、X 4 、X 5 and x 6 g / L, the corresponding 6 groups of absorbance OD were measured on the Olympus AU400 automatic biochemical analyzer 1 、OD 2 , OD 3 、OD 4 、OD 5 and OD 6 , the biochemical analyzer automatically fits a calibration curve. The calibration curve obtained on the automatic biochemical analyzer is as follows: figure 2 shown. The calibration concentration and absorbance are shown in Table 1:

[0040] Table 1 Absorbance corresponding to HP calibration concentration of...

Embodiment 2

[0053] 1. Prepare reagent R:

[0054] Reagent R contains: Tris 0.05M, polyethylene glycol 6000 2.0w / v%, Tween 20 0.2w / v%, sodium chloride 0.85w / v%, haptoglobin antibody 10.0w / v %, the pH value is 8.0.

[0055] 2. Detection: add a known concentration of C to 300 μl reagent R respectively 校准 3 μl each of calibrator and human serum sample, read the absorbance A of the reaction solution respectively 校准 and A 人血清样品 , using the automatic biochemical analyzer Olympus AU400 to automatically calculate the concentration of HP in human serum samples through the built-in single-point straight line fitting mode. The reagent of the present invention was compared with the immunoturbidimetric HP reagent purchased from Roche to measure the same patient serum, and the results are shown in Table 3:

[0056] Table 3 Reagent of the present invention compares with reference reagent measured value

[0057] sample number

Measured value of control reagent X

The measured value Y o...

Embodiment 3

[0062] A kit for detecting haptoglobin in serum, which contains reagents R1 and R2, wherein reagent R1 contains: Tris 0.05M, polyethylene glycol 6000 2.5w / v%, Tween 200.2w / v%, Sodium chloride 0.85w / v%, pH value 8.0; Reagent R2 contains: Tris 0.05M, sodium chloride 0.85w / v%, haptoglobin antibody 35w / v%, pH value 8.0.

[0063] The above kit was used to detect the content of haptoglobin in serum, and the detection method was the same as that described in Example 1.

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Abstract

The invention provides a method for simply and quickly detecting hoptoglobin in serum, namely an immunoturbidimetry, and a detecting kit thereof. The method is a detecting method designed by using the principle that the hoptoglobin is a human protein, has antigenicity and generates cohesion after combining with an antigen antibody. The hoptoglobin in the serum sample is combined with adequate specific antibody in the agent to form an insoluble immune complex; the change of the absorbance of the reaction solution is relevant to the content of the hoptoglobin in the serum sample. The method for detecting the hoptoglobin in the serum and the detecting kit in the invention have the advantages that: the serum sample is not subjected with special treatment, and can be ordinarily detected on an automatic biochemistry analyzer; the result directly reflects the true protein content of the hoptoglobin in the serum sample; therefore, the invention is used for clinic detection.

Description

technical field [0001] The invention relates to an immunological detection method, in particular to an immunological detection method for haptoglobin in serum and a detection kit thereof. Background technique [0002] Haptoglobin (HP), also known as haptoglobin, is widely present in the blood and other body fluids of humans and many mammals, and its content in individuals is relatively stable. Its biosynthesis and degradation are mainly carried out in the liver. Haptoglobin has the ability to bind to free hemoglobin, and acts like a transporter of hemoglobin. A molecule of haptoglobin can bind to a molecule of free hemoglobin and form a complex, which is quickly absorbed by mononuclear-macrophages in the blood. Cells are disposed of. This process prevents the filtration of hemoglobin from the glomerulus and avoids damage to the kidneys caused by free hemoglobin. During this metabolic process, haptoglobin is also degraded, manifested as a decrease in the level of haptoglobi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/536
Inventor 夏另朝张筠
Owner BIOSINO BIO TECH & SCI
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