Detection kit for free human chorionic gonadotropin beta subunit

A chorionic gonadotropin and detection kit technology, which is applied in biological testing, material inspection products, and analysis by making materials undergo chemical reactions, can solve the problem of low sensitivity, narrow linear range, and difficulty in full automation of enzyme-linked immunoassays and other problems, to achieve the effects of good labeling stability, improved sensitivity, and short reaction time

Pending Publication Date: 2020-03-17
NINGBO AUCHEER BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The enzyme-linked immunoassay method has methodological limitations such as low sensitivity, narrow linear range, and difficulty in realizing full automation.

Method used

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  • Detection kit for free human chorionic gonadotropin beta subunit
  • Detection kit for free human chorionic gonadotropin beta subunit
  • Detection kit for free human chorionic gonadotropin beta subunit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] A free human chorionic gonadotropin beta subunit magnetic particle chemiluminescent kit, comprising

[0025] Biotin-labeled free human chorionic gonadotropin beta subunit antibody;

[0026] Streptavidin magnetic particle suspension;

[0027] Acridinium ester-labeled free human chorionic gonadotropin beta subunit antibody;

[0028] Standard;

[0029] Cleaning fluid;

[0030] Diluent;

[0031] Chemiluminescence substrate solution A;

[0032] Chemiluminescent substrate solution B;

[0033] The streptavidin magnetic particle suspension is a magnetic particle solution coated with streptavidin.

[0034] The biotin-labeled free human chorionic gonadotropin beta subunit antibody is a monoclonal antibody.

[0035] The cleaning solution is a Tris-HCl buffer solution with a concentration of 0.025 mol / L, which contains NaCl and 0.05% Tween-20 with a concentration of 0.15 mol / L.

[0036] The chemiluminescent substrate solution A consists of 1.5% H 2 o 2 and HNO at a concen...

Embodiment 2

[0045] Included in the present invention is the preparation method of streptavidin magnetic particle solution, and concrete steps are:

[0046] Dissolve 20g of magnetic fluid and 2g of polyethylene glycol in 73.68g of absolute ethanol, transfer it to a three-necked bottle with a stirrer, condenser and nitrogen inlet, and stir at 300r / min for 30min in a nitrogen atmosphere at 65°C , then heated up to 70°C, added 19.78g styrene, 2g maleic anhydride, 1.37g acrylic acid, 0.25g divinylbenzene, 6g benzoyl peroxide in sequence, after 10min, the reactant was in the form of an emulsion, and the nitrogen atmosphere was maintained. 70°C, the stirring speed is 300r / min, and the reaction time is 12h. After the reaction is completed, the polystyrene magnetic particles are collected for future use.

[0047]The PBS buffer is autoclaved, 1 mg of streptavidin is dissolved in 1 ml of PBS buffer, and 200 ul is packed in an EP tube for use. Soak 50mg of polystyrene magnetic particles in ultrapure...

Embodiment 3

[0050] Utilize the standard product to detect the kit of the present invention, and the detection results are as shown in Table 1, and the minimum detection value of sensitivity is 0.1ng / ml; the calibration curve is made according to the chemiluminescence value and the concentration, as shown in Figure 2, the linear coefficient r 2 =0.9962, the linear range is 1~150ng / ml.

[0051] Table 1

[0052]

[0053] Example 3

[0054] The present invention has also carried out repeated test, to verify the accuracy of this kit.

[0055] Getting concentration is that the free human chorionic gonadotropin beta subunit standard substance of 5ng / ml carries out 10 tests, utilizes linear equation y=1590x+5937.1, calculates detection concentration value.

[0056] The test results are shown in Table 3. The measured average value is 5.007ng / ml, the standard deviation is 0.096, and the intra-assay coefficient of variation is 1.9%. The test results of this kit are highly accurate and can be us...

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Abstract

The invention discloses a detection kit for a free human chorionic gonadotropin beta subunit. The detection kit comprises a biotin-labeled free human chorionic gonadotropin beta subunit antibody, a streptavidin magnetic particle suspension, an acridinium ester-labeled free human chorionic gonadotropin beta subunit antibody, a standard substance, a cleaning solution, a diluent, a chemiluminescent substrate solution A and a chemiluminescent substrate solution B. The streptavidin and the biotin have high-specificity binding capacity, streptavidin and biotin-labeled high-purity antibody non-covalent bonds are specifically bound, the cascade amplification effect is achieved and the reaction is highly specific. The high specificity of antibody-antigen reaction and high sensitivity of acridiniumester luminescence are combined, photons generated by reaction are captured by acridinium ester to detect the concentration of the product, and the detection kit has the characteristics of higher sensitivity, short reaction time, simple operation and high anti-interference performance.

Description

technical field [0001] The invention relates to the field of in vitro diagnosis, in particular to a detection kit for free human chorionic gonadotropin beta subunit. Background technique [0002] Human chorionic gonadotropin is a glycoprotein secreted by the trophoblast cells of the placenta. It is composed of glycoproteins of α and β dimers. The α subunit interacts with FSH (follicle stimulating hormone) and LH ( Luteinizing hormone) and TSH (thyroid-stimulating hormone) are basically similar, so they can cross-react with each other, but the structure of the β subunit is not similar, and the β subunit is used to prepare specific antibodies to measure HCG in blood. [0003] The examination of HCG is of great significance to the diagnosis of early pregnancy, and has certain value in the diagnosis, differentiation and observation of the course of diseases such as pregnancy-related diseases and trophoblastic tumors. Currently, the immunoassay methods used to detect free human ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/76G01N21/76
CPCG01N33/76G01N21/76
Inventor 周义正黄丹娣刘超波
Owner NINGBO AUCHEER BIOTECHNOLOGY CO LTD
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