A Biochemical Analysis Method Based on Magnetic Separation for Simultaneous Detection of Multiple Targets

A technology for biochemical analysis and target objects, applied in the field of biochemical analysis, can solve problems such as cumulative errors, affecting the stability and accuracy of detection results, and complicated operations, and achieves strong capabilities, improved detection sensitivity and accuracy, and fewer operating steps Effect

Active Publication Date: 2022-06-28
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method achieves ultra-sensitive and absolute quantitative analysis of disease-related proteins, but it cannot detect multiple targets at the same time, and the method is complicated to operate, requiring multiple steps to elute the detection particles from the immune complex , it is easy to cause cumulative errors, which affects the stability and accuracy of the test results

Method used

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  • A Biochemical Analysis Method Based on Magnetic Separation for Simultaneous Detection of Multiple Targets
  • A Biochemical Analysis Method Based on Magnetic Separation for Simultaneous Detection of Multiple Targets
  • A Biochemical Analysis Method Based on Magnetic Separation for Simultaneous Detection of Multiple Targets

Examples

Experimental program
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Effect test

Embodiment 1

[0047] Example 1 Quantitative detection of procalcitonin (PCT) in whole blood

[0048] (1) Preparation of nano-magnetic particle-procalcitonin capture antibody conjugate and polystyrene microsphere-procalcitonin detection antibody conjugate

[0049] Take 2 mg of carboxyl-modified magnetic nanoparticles (200 nm, 10 mg / mL), wash twice with MES buffer (pH=6.0), resuspend in MES buffer and add 50 μL EDC (10 mg / mL) and 25 μL NHS (10 mg / mL) mL), mixed and activated at room temperature for 15 min. After activation, wash twice with PBS buffer, resuspend in PBS buffer, add 0.2 mg of procalcitonin (PCT) to capture the antibody, and react at room temperature for 2-4 hours; after the reaction, block with 1% BSA solution for 30 minutes, After the blocking was completed, the cells were washed twice with PBST buffer, and the nano-magnetic particle-procalcitonin capture antibody conjugate was resuspended in PBS buffer and stored at 4°C for later use.

[0050] Take 2 mg of carboxyl-modified ...

Embodiment 2

[0068] Embodiment 2 Quantitative detection of antibiotic molecules chloramphenicol, amoxicillin and neomycin

[0069] (1) Preparation of nanomagnetic particles-antibody conjugates of different kinds of antibiotics and polystyrene microspheres of different particle sizes-complete antigen conjugates of different kinds of antibiotics

[0070] Take 3 mg of carboxyl-modified magnetic nanoparticles (200 nm, 10 mg / mL) into 3 parts, each 1 mg, wash twice with MES buffer (pH=6.0), and add 20 μL of MES buffer after resuspending each part. EDC (10 mg / mL) and 10 μL NHS (10 mg / mL) were activated at room temperature for 15 min and mixed well. After activation, they were washed twice with PBS buffer, resuspended in PBS buffer, and then added 0.1 mg of chloramphenicol antibody, 0.1 mg of amoxicillin antibody and 0.1 mg of neomycin antibody, respectively, and reacted at room temperature for 2-4 hours; After the reaction, the cells were blocked with 1% BSA solution for 30 min, washed twice wit...

Embodiment 3

[0082] Example 3 Quantitative detection of C-reactive protein (CRP), procalcitonin (PCT) and interleukin-6 (IL-6) in whole blood

[0083] (1) Preparation of nano-magnetic particles-different kinds of biomarkers capture antibody conjugates and polystyrene microspheres of different colors-different kinds of biomarkers detection antibody conjugates

[0084]Take 3 mg of carboxyl-modified magnetic nanoparticles (200 nm, 10 mg / mL) into 3 parts, each 1 mg, wash twice with MES buffer (pH=6.0), and add 20 μL of MES buffer after resuspending each part. EDC (10 mg / mL) and 10 μL NHS (10 mg / mL) were activated at room temperature for 15 min and mixed well. After activation, wash twice with PBS buffer, resuspend in PBS buffer, add 0.1 mg of CRP capture antibody, 0.1 mg of PCT capture antibody, and 0.1 mg of IL-6 capture antibody, respectively, and react at room temperature for 2-4 hours; the reaction ends After blocking with 1% BSA solution for 30 min, washed twice with PBST buffer after bl...

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Abstract

The invention discloses a biochemical analysis method for simultaneous detection of multiple targets based on magnetic separation. The method combines biochemical reactions with visualized microsphere counting, machine vision and deep learning, and uses polymer microspheres as signal probes. The biorecognition molecules corresponding to the target to be detected are coupled to the surface of the nano-magnetic particles and the signal probe, respectively, for immune reaction or DNA molecular hybridization reaction, and the signal probe in the reaction complex is directly imaged by optical microscopy after magnetic separation Count, and use signal probes of different particle sizes or colors to distinguish different detection objects, and finally realize the simultaneous detection of multiple targets.

Description

technical field [0001] The invention belongs to the field of biochemical analysis, and relates to a biochemical analysis method for simultaneously detecting multiple target substances. Background technique [0002] With the development of the country and the improvement of people's living standards, the challenges faced by food safety, in vitro diagnosis and environmental monitoring are increasing day by day. Efficient detection methods have an extremely important impact on ensuring food safety, ecological environment safety, and promoting human health. . Most of the traditional detection methods rely on large-scale precision instruments and equipment. Although they have the characteristics of high precision and good accuracy, most of the instruments are expensive and inconvenient to carry. Before use, complex pretreatment of the test samples is required. There are also stricter requirements for personnel. [0003] Immunoassays based on antibody-antigen recognition reactio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/58G01N33/533
CPCG01N33/54313G01N33/588G01N33/533G01N33/585G01N33/582
Inventor 陈翊平周阳王知龙赵维琦鲁鹏
Owner HUAZHONG AGRI UNIV
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