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Homogeneous chemiluminescence detection kit and application thereof

A homogeneous chemiluminescence and detection kit technology, applied in the field of chemiluminescence, can solve problems such as narrow detection range or linear range, and achieve the effects of reducing non-specific adsorption, improving luminous efficiency and improving accuracy

Pending Publication Date: 2020-01-31
BEYOND DIAGNOSTICS (SHANGHAI) CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although the detection sensitivity of the chemiluminescence detection method can be improved to a certain extent by adopting the method described in the art, the detection range or linear range is relatively narrow

Method used

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  • Homogeneous chemiluminescence detection kit and application thereof
  • Homogeneous chemiluminescence detection kit and application thereof
  • Homogeneous chemiluminescence detection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0199] Example 1: Preparation of homogeneous chemiluminescence detection kit

[0200] 1. Preparation of acceptor microspheres

[0201] 1. Prepare a 25 mL round bottom flask, add 0.1 g of Europium (Ⅲ) complex, 10 mL of 95% ethanol, magnetically stir, and heat the water bath to 70° C. to obtain a solution of Europium (Ⅲ) complex.

[0202] 2. Prepare a 100mL three-necked flask, add 10mL 95% ethanol, 10mL water and 10mL 10% concentration, 200nm particle size polystyrene microspheres coated with carboxydextran hydrogel, magnetically stir, and the water bath is heated to 70 ℃.

[0203] 3. Slowly add the europium (III) complex solution in step 1 dropwise to the three-necked flask in step 2, stop stirring after reacting at 70°C for 2 hours, and cool naturally to obtain an emulsion.

[0204] 4. Centrifuge the above emulsion for 1 hour at 30000g, discard the supernatant after centrifugation, and then resuspend it with 50% ethanol. After repeated centrifugation and cleaning for 3 times, the solu...

Embodiment 2

[0235] Example 2: Homogeneous chemiluminescence detection method

[0236] Add 25μL of the sample to be tested, 25μL of biotin-labeled antibody, 175μL of donor reagent, and 25μL of acceptor reagent to the test sample hole position, additional reagent hole position, first reagent hole position, and second reagent hole position of the reagent cup strip. Then, put the reagent cup strip in the POCT analyzer developed by Boyang Biotechnology (Shanghai) Co., Ltd., the sample adding mechanism takes the corresponding volume of the sample to be tested, adds it to the additional reagent hole, vibrates, and incubates at 37°C 10 minutes; add the incubated liquid in the additional reagent well to the first reagent well and shake, incubate at 37°C for 10 minutes to form a mixed liquid; continue to add the mixed liquid to the second reagent well, shake, 37 Incubate at ℃ for 10 minutes to form the mixture to be tested. Use the laser emitted by the exciter in the detection assembly to irradiate t...

Embodiment 3

[0237] Example 3: Detection of luminous signal

[0238] Using the method described in Example 2 and the kit described in Example 1 (a kit containing receptor microspheres coated with Anti-PCT antibody I and biotin-labeled Anti-PCT antibody II as an example) was used to detect the test sample The test results are shown in Table 1.

[0239] Table 1

[0240]

[0241] It can be seen from Table 1 that under the condition that the particle size of the donor microspheres is fixed, as the particle size of the acceptor microspheres increases, the detected luminescence signal value gradually decreases. When the particle diameters are equal, the detected luminescence signal value is the largest. And when the particle size of the donor microspheres and the acceptor microspheres are both 200nm, the detected luminescence signal value is the best, and the detection sensitivity is the best.

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Abstract

The invention relates to a homogeneous chemiluminescence detection kit and application thereof. The kit comprises a donor reagent and a receptor reagent, wherein the donor reagent comprises donor microspheres, the donor microspheres can generate active oxygen in an excited state, and the surfaces of the donor microspheres are coated with markers; the receptor reagent comprises receptor microspheres, the receptor microspheres can react with the active oxygen to generate detectable chemiluminescence signals, the surfaces of the receptor microspheres are coated with biomolecules, and the biomolecules can be specifically combined with target molecules to be detected; the particle size of the receptor microspheres is equal to that of the donor microspheres. The particle size of the donor microspheres in the kit is equal to that of the receptor microspheres, and the precision and sensitivity of the kit are improved.

Description

Technical field [0001] The invention belongs to the technical field of chemiluminescence, and specifically relates to a homogeneous chemiluminescence detection kit and an application thereof. Background technique [0002] So far, the methods for the determination of trace biologically active substances in organisms have gone through various stages including radioimmunoassay, analysis (RIA), fluorescence immunoassay (FIA), enzyme-linked immunoassay (EIA) and chemiluminescence analysis. This evolutionary process is mainly based on the continuous improvement of the sensitivity, accuracy, and simplicity of the detection method. [0003] Chemiluminescence analysis is a detection method using light waves emitted by chemiluminescent substances. Chemiluminescent substances are used as labels in nucleic acid detection and immunoassay. For example, a certain molecule in a specific binding pair can be combined with a luminescent substance through a variety of ways to form a luminescent comp...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N21/01
CPCG01N21/76G01N21/01G01N33/54313
Inventor 杨阳赵卫国刘宇卉李临
Owner BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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