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Kit for detecting estradiol by utilizing magnetic particle chemiluminescence immunoassay

A chemiluminescence immunity and chemiluminescence technology, applied in the field of immunoassay, can solve problems affecting detection sensitivity and accuracy, and achieve good market application prospects, high sensitivity, and easy to use

Inactive Publication Date: 2010-02-03
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the actual immunoassay, due to the large amount of impurities contained in the sample to be tested, the detection sensitivity and accuracy are affected to a certain extent.

Method used

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  • Kit for detecting estradiol by utilizing magnetic particle chemiluminescence immunoassay
  • Kit for detecting estradiol by utilizing magnetic particle chemiluminescence immunoassay
  • Kit for detecting estradiol by utilizing magnetic particle chemiluminescence immunoassay

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1 Preparation I of kit of the present invention

[0051] 1), preparation of estradiol series calibrator

[0052] A) Preparation of hormone-free human serum:

[0053] Put 60mL of normal mixed human serum into four volumetric flasks in equal parts, then add 8.0g of activated carbon respectively, mix evenly with a vortex mixer by inversion, shake for 8h, centrifuge at 6000rpm for 20min, filter the supernatant, add Preservative Proclin-300 with a concentration of 0.5‰ by volume, stored in a frozen state.

[0054] B) Preparation of estradiol calibrator:

[0055] E 2 Dilute the pure product into a calibrator, aliquot E 2 A total of 6 bottles of calibrators with pure concentrations of 0, 15, 50, 150, 500, and 1000 pg / mL.

[0056] 2) Preparation of goat anti-rabbit polyclonal antibody-coated magnetic particle solution

[0057] The goat anti-rabbit polyclonal antibody-coated magnetic particle solution was prepared by glutaraldehyde two-step method. The specific ...

Embodiment 2

[0098] Embodiment 2 Preparation II of the kit of the present invention

[0099] 1), the preparation of estradiol series calibrator, with embodiment 1

[0100] 2) Preparation of goat anti-rabbit polyclonal antibody-coated magnetic particle solution

[0101] The goat anti-rabbit polyclonal antibody-coated magnetic particle solution was prepared by glutaraldehyde two-step method. The specific operation steps were as follows: Activate the magnetic particle with a particle size of 2.0 μm with glutaraldehyde, stir at room temperature, and mix for 2 hours. , add a magnetic field with a magnetic field strength of 2000 gauss, let stand for 25 minutes, pour out the supernatant, wash three times with 0.01mol / L phosphate buffer saline with a pH value of 7.4, and suspend with the buffer, the concentration is 50mg / mL; Add 80 μg of purified goat anti-rabbit polyclonal antibody to the suspension in milliliters, stir overnight at 4°C, add a magnetic field, wherein the magnetic field strength ...

Embodiment 3

[0138] Embodiment 3 Preparation III of the kit of the present invention

[0139] 1), the preparation of estradiol series calibrator, with embodiment 1

[0140] 2) Preparation of goat anti-rabbit polyclonal antibody-coated magnetic particle solution

[0141] The goat anti-rabbit polyclonal antibody-coated magnetic particle solution was prepared by glutaraldehyde two-step method. The specific operation steps were as follows: Activate the magnetic particle with a particle size of 3.0 m with glutaraldehyde, stir at room temperature, and mix for 2 hours. , add a magnetic field with a magnetic field strength of 2000 gauss, let stand for 20 minutes, pour out the supernatant, wash three times with 0.01mol / L phosphate buffer saline with a pH value of 7.4, and use the buffer to suspend at a concentration of 75 mg / mL; Add 60 μg of purified goat anti-rabbit polyclonal antibody to the suspension in milliliters, stir overnight at 4°C, add a magnetic field, wherein the magnetic field streng...

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Abstract

The invention discloses a kit for detecting the estradiol (E2) by utilizing magnetic particle chemiluminescence immunoassay. The kit comprises estradiol series calibrator, magnetic particle solution enveloped by the second antibody, estradiol marked by horse radish peroxidase, estradiol antibody, chemiluminescence substrate liquid and concentrated washing liquid. The invention further discloses apreparation method of the kit. The kit of the invention can quantitively detect the content of estradiol in the serum and blood plasma samples of human body, and can simultaneously detect a large number of samples, thus having advantages of simpleness, convenience, rapidness, stability, high sensitivity and the like, and providing a extremely valuable detection method for clinical diagnosis and scientific research.

Description

technical field [0001] The invention belongs to the field of immune analysis, and in particular relates to a kit for detecting estradiol by using magnetic particle chemiluminescence immune technology and a preparation method thereof. The kit of the invention combines the immunomagnetic particle separation technology and the chemiluminescence immunoassay technology. Background technique [0002] Estradiol (E 2 ) is the most biologically active female hormone, which is mainly synthesized by the ovary and a small amount is secreted by the adrenal gland. influences. Serum estradiol concentration is one of the functional indicators of the hypothalamus-pituitary-gonadal axis, which has certain value and significance in the diagnosis of certain endocrine and gynecological diseases. Changes in estradiol levels can manifest different lesions. Elevated estradiol can be seen in female precocious puberty, twin pregnancy or multiple pregnancy, ovarian sex cord stromal cell tumor, end...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N21/76G01N33/535
Inventor 林金明辛天兵肖勤
Owner TSINGHUA UNIV
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