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Dissociate human chorionic gonadotrophin beta-subunit magnetic particle chemiluminescence quantitative assay kit and its preparation method

A chorionic gonadotropin and chemiluminescence technology, applied in the field of immunoassay medicine, can solve the problems of limited popularization and use, inability to be widely used in clinical diagnosis and scientific research, and achieve no radioactive contamination, simple, reliable, and specific sample pretreatment process. strong effect

Inactive Publication Date: 2013-05-01
BEIJING LEADMAN BIOCHEM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the chemiluminescence immunoassay kits in the prior art are imported closed automatic chemiluminescence detection systems, which require expensive automatic chemiluminescence detectors, thus limiting the promotion and use, and cannot be widely used in clinical diagnosis and scientific research

Method used

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  • Dissociate human chorionic gonadotrophin beta-subunit magnetic particle chemiluminescence quantitative assay kit and its preparation method
  • Dissociate human chorionic gonadotrophin beta-subunit magnetic particle chemiluminescence quantitative assay kit and its preparation method
  • Dissociate human chorionic gonadotrophin beta-subunit magnetic particle chemiluminescence quantitative assay kit and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Preparation of the kit for quantitative determination of free human chorionic gonadotropin β subunit magnetic particle chemiluminescence method of the present invention

[0041] 1. Preparation of F-hCGβ calibrator

[0042] The pure F-hCGβ was diluted with newborn bovine serum into a calibrator, and divided into 6 bottles of 0 ng / mL, 5 ng / mL, 15 ng / mL, 50 ng / mL, 100 ng / mL, and 200 ng / mL.

[0043] 2. Preparation of magnetic particle solution coated with anti-FITC polyclonal antibody

[0044] Apply a magnetic field to magnetic particles with a particle size of 1 μm, let them stand for 15 minutes, pour out the supernatant, wash 3 times with 25mM MES buffer solution with pH=4.7, and suspend with the buffer solution, the concentration is 50mg / mL; Add 2 mg of anti-FITC polyclonal antibody to the suspension, and mix well at room temperature; prepare an EDC solution with a concentration of 10 mg / mL with deionized water, and add 1 mL of EDC solution with a concentratio...

Embodiment 2

[0054] Embodiment 2 The usage method of the kit of the present invention

[0055] 1. Sample requirements

[0056] Take 5.0ml of venous blood into a glass test tube without adding anticoagulant, let it stand at room temperature, and then separate the serum part by centrifugation (3000rpm×5min).

[0057] 2. Detection method

[0058] Reagents should be equilibrated to room temperature after being taken out from storage conditions before being used for detection; Magnetic separation reagents: Mix thoroughly before use to ensure that the magnetic particles are evenly suspended, and cannot be stirred with a magnetic stirrer; Cleaning concentrate: Dilute 15 times with deionized water , mix well; set up a 37°C water bath; please put the chemiluminescence detector in the standby state; prepare and mark the test tubes as needed.

[0059] The specific operation steps of using this kit to carry out the experiment according to the method of implementation 2 are as follows:

[0060] Add ...

Embodiment 3

[0061] Embodiment 3 The methodological test of the kit of the present invention

[0062] The test kit prepared in Example 1 is tested according to the conventional manufacturing and testing procedures in the art, and the results are as follows:

[0063] 1. Determination of kit precision

[0064] (1) Analytical internal precision

[0065] A batch of the kit prepared in Example 1 was used to measure three different concentrations of serum, namely low, medium and high, and 10 wells were measured in parallel, and the intra-assay coefficient of variation was 4.35% to 6.25%.

[0066] Table 1 Analytical Inner Precision Test

[0067] Determination of serum concentration (ng / mL) Measurement times Intra-analytical CV(%) 10.58 10 6.25 78.65 10 3.69 126.36 10 4.35

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Abstract

The invention relates to the medical field of immunoassay, and concretely provides a dissociate human chorionic gonadotrophin beta-subunit magnetic particle chemiluminescence quantitative assay kit having the advantages of simplicity, rapidness, high sensitivity, wide linear range and good stability, and its preparation method. The method combines a magnetic particle immune separating technology to apply an enzymatic chemiluminescence substrate on the basis of enzyme-linked immunosorbent assay, and allows an optical signal generated by the detection of the luminescence substrate to substitute a chromogenic substrate in enzyme immune assay, so the method has the advantages of substantially improved sensitivity, simple operation and wide practicality, can be applied to open semi-automatic chemiluminescence detectors, can also be applied to full-automatic measure systems, and can realize batch and fast detection, low use cost, and easy popularization and application.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine, and specifically provides a simple, rapid, high-sensitivity, wide linear range and good stability free human chorionic gonadotropin beta subunit magnetic particle chemiluminescence quantitative assay kit and its Preparation Background technique [0002] hCG is a glycoprotein produced by the syncytiotrophoblast of the placental chorion, which is composed of α and β units bound together by ionic and hydrophobic bonds. The α subunit is shared by the anterior pituitary hormones, and it is similar in structure to the α subunits of FSH, LH, and TSH. The structure of the β subunit is different from the β subunit of LH, etc., and can be distinguished in terms of immunological activity. Most of the β subunits in the blood exist in the form of whole molecules, and the free β subunits (F-hCGβ) account for about 1% to 5% of hCG. The free β subunit in the blood is produced 6 to 8 days after concepti...

Claims

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Application Information

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IPC IPC(8): G01N33/76G01N21/76
Inventor 不公告发明人
Owner BEIJING LEADMAN BIOCHEM
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