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Kit for detecting hyaluronic acid and detection method and application of kit

A technology of hyaluronic acid and kits, applied in biological testing, measuring devices, material inspection products, etc., can solve problems such as radioactive pollution, long detection cycle, and unsatisfactory repeatability of ELISA methods

Inactive Publication Date: 2015-06-10
SHENZHEN NEW INDS BIOMEDICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the main laboratory analysis methods for serum hyaluronic acid include RIA (radioimmunoassay) and ELISA (enzyme-linked immunosorbent assay). The shortcoming of long cycle, and the repeatability of ELISA method is not ideal

Method used

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  • Kit for detecting hyaluronic acid and detection method and application of kit
  • Kit for detecting hyaluronic acid and detection method and application of kit
  • Kit for detecting hyaluronic acid and detection method and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] A test kit for detecting hyaluronic acid, comprising the following components:

[0064] 1) Magnetic microsphere system: magnetic microsphere solution coated with hyaluronic acid antigen (HA antigen).

[0065] Among them, the working concentration of the magnetic microspheres: 0.5 mg / ml, and the working concentration of the HA antigen: 2 μg / ml.

[0066] 2) Marker system: ABEI solution labeled with anti-hyaluronan-binding protein monoclonal antibody (HABP monoclonal antibody).

[0067] Wherein: the working concentration of anti-hyaluronan-binding protein monoclonal antibody: 25 μg / ml, and the working concentration of ABEI: 100 ng / ml.

[0068] 3) Free hyaluronic acid binding protein solution.

[0069] Wherein: the working concentration of hyaluronic acid binding protein: 600ng / ml.

[0070] 4) Calibrator solution: a low-point calibrator solution with a hyaluronic acid concentration of 35.566 ng / ml and a high-point calibrator solution with a concentration of 1124.68 ng / ml...

Embodiment 2

[0096] A test kit for detecting hyaluronic acid, which is the same as the test kit in Example 1.

[0097] The method for detecting hyaluronic acid (one-time cleaning method) of the present embodiment comprises the following steps:

[0098] 1) Reaction: Add 40 μl of the sample to be tested (serum), high and low concentration calibrator into the cuvette respectively, then add 20 μl of magnetic microsphere solution coated with HA antigen, and add 100 μl of free hyaluronic acid binding protein solution at the same time And 100 μl of HABP monoclonal antibody-labeled ABEI solution, incubate at 37°C for 15 minutes, so that the HA in the sample to be tested competes with the HA antigen attached to the magnetic microspheres for binding to the hyaluronic acid binding protein.

[0099] 2) Detection: Apply an external magnetic field to precipitate the above reaction product, remove the supernatant, wash with buffer, and then add chemiluminescent exciters (NaOH and H 2 o 2 ), detect the ...

Embodiment 3

[0101] A test kit for detecting hyaluronic acid, which is basically the same as the test kit in Example 1, except that:

[0102] 1) Magnetic microsphere system:

[0103] A solution of magnetic microspheres coated with streptavidin (SA).

[0104] Among them, the working concentration of the magnetic microspheres: 0.5 mg / ml, and the working concentration of the SA antigen: 1 μg / ml.

[0105] Biotin-labeled hyaluronic acid antigen solution.

[0106] Among them, the working concentration of Biotin: 200ng / ml, the working concentration of HA antigen: 2000ng / ml.

[0107] The preparation method of the kit for detecting hyaluronic acid in this example refers to the preparation method in Example 1, except for the following steps, the rest are the same as the method in Example 1.

[0108] 1. Preparation of magnetic microsphere system.

[0109] 1. Preparation of magnetic microsphere solution coated with streptavidin (SA).

[0110] The preparation method of the magnetic microsphere sol...

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Abstract

The invention discloses a kit for detecting hyaluronic acid and a detection method and application of the kit, and belongs to the technical field of in-vitro diagnosis detection. The kit comprises the following components: (1) a magnetic microsphere system comprising magnetic microspheres which are directly or indirectly connected with a first binding agent, and (2) a marker system comprising a second binding agent which is directly or indirectly connected with a mark tracer. According to the kit and the detection method disclosed by the invention, the hyaluronic acid is detected by adopting chemiluminescence immunoassay, a chemiluminescence determination technology with high sensitivity is combined with high-specificity immunoreaction, and the kit has the characteristics of high sensitivity and high accuracy.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis and detection, in particular to a kit for detecting hyaluronic acid and a detection method and application thereof. Background technique [0002] Liver fibrosis is a common pathway of liver damage caused by various chronic liver diseases. Serological markers of liver fibrosis include direct markers that reflect extracellular matrix production and indirect markers that reflect liver synthesis, metabolism, and reserve functions. Various comprehensive serological marker evaluation models can determine significant fibrosis and cirrhosis, reduce unnecessary liver biopsy, and can assist in the diagnosis and staging of diseases, prognosis assessment, and treatment monitoring. [0003] HA (hyaluronic acid) is a substance composed of mucopolysaccharides, commonly found in the matrix of connective tissue, its molecular structure is a polymer of disaccharides, the basic units are β-glucuronic aci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/543G01N33/54326G01N33/6893G01N2800/085
Inventor 饶微李武袁锦云唐富强林冬霞李婷华
Owner SHENZHEN NEW INDS BIOMEDICAL ENG
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