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Method for detecting human chorionic gonadotropin beta through flash type homogeneous chemiluminescence

A chorionic gonadotropic and homogeneous chemiluminescence technology, which is applied in the biological field, can solve the problems of cumbersome operation and long detection time, and achieve the effects of good repeatability, less time and less antibody consumption

Pending Publication Date: 2019-07-05
杭州普鲁米生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This method uses magnetic particles as a solid support, and requires an external magnetic field and washing steps during the detection process, so it has the disadvantages of cumbersome operation and long detection time

Method used

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  • Method for detecting human chorionic gonadotropin beta through flash type homogeneous chemiluminescence
  • Method for detecting human chorionic gonadotropin beta through flash type homogeneous chemiluminescence
  • Method for detecting human chorionic gonadotropin beta through flash type homogeneous chemiluminescence

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Embodiment 1

[0061] Example 1. A method for detecting human chorionic gonadotropin beta by flash-type homogeneous chemiluminescence technology

[0062] The β-HCG monoclonal antibody Ⅰ is coupled with the luminescent substrate 9,10-dihydroacridine, and the β-HCG monoclonal antibody Ⅱ is coupled with horseradish peroxidase for the detection of human by flash-type homogeneous chemiluminescent method. Chorionic gonadotropin beta; the following steps are performed in sequence:

[0063] 1), β-HCG monoclonal antibody I coupled with 9,10-dihydroacridine:

[0064] Dissolve 1mg of 9,10-dihydroacridine (luminescence substrate) in 2ml of DMF to obtain 9,10-dihydroacridine solution;

[0065] Take 100 ul of 9,10-acridine solution and add 100 ul of β-HCG monoclonal antibody I with a concentration of 10 mg / ml, then add 800 ul of 0.05M boric acid buffer (PH8.8), and rotate for 1 hour at room temperature.

[0066] Add the obtained reaction solution into a dialysis bag with a molecular weight cut-off of 70...

Embodiment 2

[0088] Embodiment 2, a method for detecting human chorionic gonadotropin β by a flash-type homogeneous chemiluminescent technique:

[0089] Goat anti-mouse IgG secondary antibody coupled with luminescent substrate 9,10-dihydroacridine, goat anti-rabbit IgG secondary antibody coupled with horseradish peroxidase, used for flash-type homogeneous chemiluminescent detection of human chorion Gonadotropin beta. Follow the steps in order:

[0090] 1), goat anti-mouse IgG secondary antibody coupled to 9,10-dihydroacridine:

[0091] Dissolve 1mg of the luminescent substrate 9,10-acridine in 2ml of DMF to obtain a 9,10-acridine solution;

[0092] Take 100 ul of 9,10-acridine solution and add 100 ul of goat anti-mouse IgG secondary antibody with a concentration of 10 mg / ml, then add 800 ul of 0.05M boric acid buffer (PH8.8), and rotate at room temperature for 1 hour.

[0093]Add the obtained reaction solution into a dialysis bag with a molecular weight cut-off of 7000, place it in PBS ...

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Abstract

The invention discloses a method for detecting human chorionic gonadotropin beta through flash type homogeneous chemiluminescence. The method comprises the following steps: taking serum as a to-be-detected sample; enabling antibody I to couple with chemiluminescent substrate to obtain antibody I-A; enabling the antibody II to couple with horse radish peroxidase to obtain antibody II-HRP; designingtwo detection systems, adding excitation liquor after incubating the detection systems, and detecting a luminescent signal; performing gradient dilution on the beta-HCG sample, taking the gradient-diluted beta-HCG sample as a specimen to detect, thereby acquiring the luminescent signal and the formula corresponding to the human chorionic gonadotropin beta concentration; taking the to-be-detectedsample as the specimen to detect, thereby finally obtaining the concentration of the human chorionic gonadotropin beta in the to-be-detected sample. The method for detecting the human chorionic gonadotropin beta disclosed by the invention has the features of being fast, and high in sensitive.

Description

technical field [0001] The invention belongs to the field of biology, and in particular relates to a method for detecting human chorionic gonadotropin beta by a flash type homogeneous chemiluminescence method. Background technique [0002] Human chorionic gonadotropin β (β-HCG), as the β subunit of human chorionic gonadotropin (HCG), is a glycoprotein hormone secreted by the trophoblast cells of the placenta during pregnancy. β-HCG contains 145 amino acids, which is specific to HCG and determines the biological activity of glycoproteins. In the case of non-pregnancy, the HCG content in the body is very low, and it proliferates rapidly in the first 8 weeks of pregnancy to maintain pregnancy. Therefore, detecting the HCG content in women's urine or blood in early pregnancy is of great significance for the diagnosis of early pregnancy and pregnancy-related diseases. At present, studies have found that β-HCG is expressed at different levels in various tumors, such as cervical ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/76
CPCG01N33/76G01N2333/59
Inventor 江学成杨晓光
Owner 杭州普鲁米生物科技有限公司
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