Immune analysis method for determining estriol content

A technology of estriol and analytical method, which is applied in the field of marker immunoassay, can solve the problems of complex maintenance and management, difficult repair, high price and impossible

Inactive Publication Date: 2012-05-16
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this imported instrument is expensive and cannot be used in conjunction with domestic instruments, and the maintenance and management are complicated. Once damaged, it is not easy to repair, which greatly limits its application.

Method used

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  • Immune analysis method for determining estriol content
  • Immune analysis method for determining estriol content
  • Immune analysis method for determining estriol content

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] This example is to use the magnetic microparticles coated with estriol antibody of the present invention in the detection of estriol radioimmunoassay.

[0031] First, before the reaction, the magnetic microparticles coated with anti-E3 were diluted with a diluent at a ratio of 1:6.

[0032] Add E3 standard (or analyte), 100 μl of magnetic microparticles and 50 μl to the reaction test tube in sequence 125 I-E3 (35000cpm), shaking reaction at 37°C for 45min. After the reaction, the supernatant was decanted on a magnetic separator and washed 2 to 3 times, each time adding 300 μl of washing solution, which was PBST buffer solution (pH 7.4). The supernatant was discarded, and the radioactive count was measured on the DFM-96 multi-tube automatic gamma counter produced by Anhui Hefei Zhongcheng Electromechanical Technology Co., Ltd., and the logit-to-log data equation processing system was used to measure, and the relevant parameters and the concentration of the sample to be ...

Embodiment 2

[0039] This example is the use of the magnetic microparticles coated with estriol antibody of the present invention in the detection of estriol by chemiluminescence immunoassay.

[0040] First, before the reaction, the magnetic microparticles coated with anti-E3 were diluted with a diluent at a ratio of 1:6.

[0041] Add 100 μl each of E3 standard (or analyte), E3-HRP, and anti-E3-coated magnetic microparticles to the reaction test tube in sequence, and shake and react at 37°C for 60 minutes. After the reaction, the supernatant was decanted on a magnetic separator and washed 2 to 3 times, each time adding 300 μl of washing solution, which was PBST buffer solution (pH 7.4). Discard the supernatant, add 300 μl of chemiluminescence substrate solution, and place it in the dark at 37°C for 20 minutes, then detect the chemiluminescence count value on the BHP9504 microplate luminescence analyzer produced by Hamamatsu, Japan, and process according to the Logit versus log data equation...

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Abstract

The invention provides a method for coating estriol antibodies on the surfaces of magnetic corpuscules for the estriol immunoassay detection. The method is technically characterized in that the antibodies of the antigen to be detected are conjugated with the surfaces of magnetic corpuscules via covalent bonds. The magnetic corpuscules are completely washed before being coated, the magnetic corpuscules and the estriol antibody serum are further added to a specified buffer solution system, and react while vibrating at the room temperature. The estriol antibody-coated magnetic corpuscules can bedirectly involved in an immunoreaction so as to be applied in the immunoassay detection of the estriol after being enclosed. During the processes of the antibody coating and the assay measurement, a magnetic separator can be combined to carry out the precipitation separation. The method has the advantages of convenient operation and low cost. The process of the reaction can be completed in one step without large valuable instruments, and the estriol antibodies can be combined with various detection instruments, and are suitable for generalization.

Description

technical field [0001] The invention is used in the field of labeling immunoassay, and is especially suitable for measuring the content of estriol by the competition method principle in the labeling immunoassay. Background technique [0002] As one of the three information transmission systems of the human body, the endocrine system plays an important role in regulating various functions of the body. The disturbance of the endocrine system will lead to the destruction of the stability of the whole internal environment of the body. As an important natural product of endocrine glands, hormone plays an irreplaceable role in regulating body functions and maintaining internal environment stability. Hormones are transmitted in low concentrations in the blood and bind to target cell receptors to regulate body functions. Too little hormone can cause some tissue or organ dysfunction, even life-threatening; too much hormone can cause endocrine hyperfunction. As one of the main smal...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/74G01N33/546
Inventor 林金明吴丹凝王栩
Owner TSINGHUA UNIV
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