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Method for detecting aggregation-induced emission combined with immunomagnetic beads and kit thereof

A technology of aggregation-induced luminescence and immunomagnetic beads, which is applied in the direction of analysis, measurement device, fluorescence/phosphorescence, etc. through chemical reaction of materials, which can solve the problems of many detection steps, difficulty in multi-index joint inspection, and environmental pollution by cleaning waste liquid. , to shorten the detection time, improve the detection sensitivity and accuracy, and omit the washing step.

Pending Publication Date: 2021-02-12
GUANGZHOU BIOKEY HEALTH TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to introduce the aggregation-induced luminescent material as a signal unit into the immune detection system, which solves the difficulties of multi-index joint detection in the existing detection system, many detection steps, and the pollution of the environment by cleaning waste liquid

Method used

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  • Method for detecting aggregation-induced emission combined with immunomagnetic beads and kit thereof
  • Method for detecting aggregation-induced emission combined with immunomagnetic beads and kit thereof
  • Method for detecting aggregation-induced emission combined with immunomagnetic beads and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Quantitative detection of C-reactive protein (CRP)

[0038] This example takes TPE-COOH aggregation-induced fluorescent microspheres as an example, and the specific operations are as follows:

[0039] The C-reactive protein standard was diluted to 0 ng / ml, 0.52 ng / ml, 1.42 ng / ml, 2.84 ng / ml, 35.7 ng / ml, 188.35 ng / ml, 339.5 ng / ml.

[0040] The labeled value of the sample to be tested is 22.35ng / ml.

[0041] 1. Aggregation-induced fluorescent microspheres label antigens or antibodies to obtain fluorescent markers

[0042] 1.1 Labeled antibody / antigen pretreatment: Take the C-reactive protein primary antibody and add it to a 30KD ultrafiltration centrifuge tube, and centrifuge at 10000r / min at 4°C for 9min to 10min. Repeat washing with binding buffer 3 to 5 times, and concentrate to a certain concentration (such as 1 mg / ml). The binding buffer is 0.05M Tris-HCl buffer at pH 8.0-9.0.

[0043] 1.2 Aggregation-induced fluorescent microspheres labeled with anti...

Embodiment 2

[0054] Example 2 Quantitative detection of procalcitonin (PCT)

[0055] This example takes TPE-CHO aggregation-induced fluorescent microspheres as an example, and the specific operations are as follows:

[0056] The procalcitonin standard was diluted to 0 ng / ml, 0.02 ng / ml, 0.5 ng / ml, 2 ng / ml, 10 ng / ml, 50 ng / ml, 100 ng / ml.

[0057] The labeled value of the sample to be tested is 0.452ng / ml.

[0058] 1. Aggregation-induced fluorescent microspheres label antibodies to obtain fluorescent markers

[0059] 1.1 Labeled antibody pretreatment: Take procalcitonin primary antibody and add it into a 30KD ultrafiltration centrifuge tube, and centrifuge at 10000r / min at 4°C for 9min to 10min. Repeat washing with binding buffer 3 to 5 times, and concentrate to a certain concentration (such as 1 mg / ml). The binding buffer is 0.05M Tris-HCl buffer at pH 8.0-9.0.

[0060] 1.2 Aggregation-induced fluorescent microsphere-labeled antibody: Wash aggregation-induced fluorescent microspheres ...

Embodiment 3

[0071] Example 3 Dual-label quantitative detection of C-reactive protein (CRP) and serum amyloid A (SAA)

[0072] In order to achieve the purpose of detection, this embodiment adopts TPE-2SO4 with similar wavelengths of excitation light and different wavelengths of emission light using aggregation-induced fluorescent materials. 3 Na + and TPE-COOH two aggregation-induced fluorescent microspheres as examples to label C-reactive protein (CRP) and serum amyloid A (SAA) respectively, the specific operations are as follows:

[0073] The C-reactive protein standard was diluted to 0 ng / ml, 0.52 ng / ml, 1.42 ng / ml, 2.84 ng / ml, 35.7 ng / ml, 188.35 ng / ml, 339.5 ng / ml.

[0074] The serum amyloid standard was diluted to 0 ng / ml, 0.93 ng / ml, 3.57 ng / ml, 15.45 ng / ml, 73.39 ng / ml, 140.19 ng / ml, 282.82 ng / ml.

[0075] The sample to be tested contains both C-reactive protein and serum amyloid, and the marked value of C-reactive protein is 10.05 ng / ml, and the marked value of serum amyloid is...

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Abstract

The invention provides a method for detecting aggregation-induced emission combined immunomagnetic beads. The method comprises the following steps: S1, respectively marking aggregation-induced fluorescent microspheres Y1, Y2... Yn by using primary antibodies of target objects X1, X2... Xn to be detected; S2, respectively coating immunomagnetic beads Z1, Z2... Zn with secondary antibodies of targetobjects X1, X2... Xn to be detected; S3, jointly incubating the fluorescent microspheres obtained in the step S1, the immunomagnetic beads obtained in the step S2 and a detection sample, and determining the content of a to-be-detected object according to a collected fluorescence value. The detection method disclosed by the invention not only can omit the cleaning of uncombined antigen / antibody, reduce the environmental pollution of cleaning waste liquid and greatly shorten the detection time, but also can improve the detection sensitivity and accuracy. In addition, due to the aggregation-induced emission molecules, aggregation-induced emission materials with different emission light can be obtained by designing different molecular structures according to detection requirements, so that multi-marker joint detection in the same detection system is more convenient to realize, the detection efficiency is higher, and the result is more accurate.

Description

technical field [0001] The invention belongs to the field of immunodetection, and in particular relates to a detection method using aggregation-induced luminescence combined with immunomagnetic beads and a kit thereof. Background technique [0002] There is a common problem in the use of fluorescent probes: when the fluorescent functional groups are aggregated, the fluorescence will be weakened or quenched, which will lead to a significant decrease or disappearance of the emitted light signal intensity of the fluorescent molecules bound to the biomolecules. In the current practice, first, in order to avoid the aggregation of fluorescent probes, its concentration and the number of probes bound to biomolecules must be strictly controlled during use, resulting in low concentration of fluorescent groups and a large drop in fluorescence intensity, making detection difficult. . The second is to use other ions such as rare earth elements as fluorescent probes, but it is necessary ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/533G01N21/64G01N21/78
CPCG01N33/54346G01N33/54326G01N33/533G01N21/6428G01N21/78
Inventor 李爱霞李根平陈洵
Owner GUANGZHOU BIOKEY HEALTH TECH CO LTD
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