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Single-pulse detection method for immune reaction with metal ion labeling

An immune reaction and metal ion technology, applied in the field of medical testing, can solve the problems of cumbersome operation of enzyme-linked immunoassay and chemiluminescence method, large batch-to-batch variation of fluorescent immunochromatography, and poor stability of markers, etc., to improve system detection Sensitivity, stable detection results, and high sensitivity

Active Publication Date: 2018-11-20
TIANJIN SAVANT BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, radioimmunoassay has radioactive pollution, which is harmful to the human body; enzyme-linked immunoassay and chemiluminescent method are cumbersome to operate and have a long detection cycle; the electrode equipment used in the electrochemical detection system and the sophisticated signal analysis system are expensive. ; Colloidal gold method, fluorescence immunochromatography method has large batch-to-batch variability and poor stability of markers; microfluidic chip method has not yet achieved commercial application

Method used

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  • Single-pulse detection method for immune reaction with metal ion labeling
  • Single-pulse detection method for immune reaction with metal ion labeling
  • Single-pulse detection method for immune reaction with metal ion labeling

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1 (no washing single pulse competition method)

[0056] 1. The preparation process of Pt nanoparticles;

[0057] 0.15g polyvinylpyrrolidone (PVP), 33.9mg chloroplatinic acid (H 2 PtCl 6 .6H 2 (2), 20mL ultrapure water, and 20mL ethylene glycol were added to the reaction vessel, and at a speed of 240r / min, the reaction was refluxed at 60°C for 3h to obtain a brown-black colloidal solution of Pt nanoparticles.

[0058] 2. Streptavidin-labeled SiO 2 Microspheres

[0059] Take 2mL of 2.5% w / v SiO 2 Microspheres (particle size 1 μm, Xi’an Ruixi Biotechnology Co., Ltd.) were washed twice with PBS buffer solution (pH=7.4) and then resuspended in 10 mL of PBS buffer solution; continued in the washed magnetic microsphere solution Add 5mg EDC and 5mg NHS, activate for 1h under stirring; wash with PBS buffer solution and resuspend in 10mL PBS buffer solution, continue to add blocking solution for blocking for 30min, wash with PBS buffer solution and resuspend in 1...

Embodiment 2

[0072] Embodiment 2 (washing single pulse competition method)

[0073] 1. Same as the process of "preparation of Pt nanoparticles" of 1 in Example 1;

[0074] 2. "streptavidin-labeled SiO 2 Microspheres" process is the same;

[0075] 3. Biotin labeling a strain of mouse anti-human FT 3 Monoclonal antibodies:

[0076] A strain of mouse anti-human FT 3 The monoclonal antibody was diluted to 1 mg / mL, and then dialyzed with sodium carbonate buffer at room temperature (25°C±5°C) and stirred in the dark for 4 hours; then 6-aminocaproic acid- N-hydroxysuccinimide-biotin (BCNHS) was formulated at 1mg / mL; in 1mL a strain of mouse anti-human FT 3Add 125 μL to 66.7 μL of the above DMF solution to the monoclonal antibody solution, mix in a glass bottle, and stir at room temperature (25°C±5°C) in the dark for 2 hours; add 9.6 μL of 1mol / L ammonium chloride solution, °C) in the dark and stirred for 10 minutes; then the mixed solution was transferred to a dialysis bag and dialyzed over...

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Abstract

The invention discloses a single-pulse detection method for an immune reaction with metal ion labeling. Single-pulse signal detection is achieved by utilizing inductively coupled plasma or microwave plasma mass spectrometry analysis technology, the immune reaction is labeled by employing metal nanoparticles, and pulse signals of metal ions of the labeled immune reaction are further detected by utilizing the inductively coupled plasma or microwave plasma mass spectrometry analysis technology, thereby achieving a purpose of indirectly detecting a to-be-detected object.

Description

technical field [0001] The invention belongs to the field of medical testing, but is not limited to this field, and specifically relates to a metal ion-labeled immunoreaction single-pulse detection method. Background technique [0002] At present, in terms of immunodiagnosis, radioimmunoassay, enzyme-linked immunoassay, chemiluminescence, and electrochemiluminescence are commonly used clinically; at the same time, in the field of rapid testing, colloidal gold method, fluorescent immunochromatography and emerging methods are commonly used. Microfluidic chip detection methods, etc. However, radioimmunoassay has radioactive pollution, which is harmful to the human body; enzyme-linked immunoassay and chemiluminescent method are cumbersome to operate and have a long detection cycle; the electrode equipment used in the electrochemical detection system and the sophisticated signal analysis system are expensive. ; The colloidal gold method and the fluorescent immunochromatography m...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N27/62G01N27/64
CPCG01N27/626G01N27/64G01N33/5302
Inventor 林斯
Owner TIANJIN SAVANT BIOTECH CO LTD
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