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Double-enhanced chemiluminescence immunoassay based on metal-enhanced luminescence and nanoparticle labeling amplification

A technology of metal nanoparticles and metal reinforcement, which is applied in the direction of chemical reaction of materials for analysis, chemiluminescence/bioluminescence, and analysis of materials, which can solve the problems of insufficient sensitivity of detection kits and achieve fast detection speed and high sensitivity Effect

Active Publication Date: 2016-06-08
GETEIN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Another purpose of the present invention is to develop different immunological detection kits based on this technology platform to solve the problem of insufficient sensitivity of existing CLIA-based detection kits

Method used

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  • Double-enhanced chemiluminescence immunoassay based on metal-enhanced luminescence and nanoparticle labeling amplification
  • Double-enhanced chemiluminescence immunoassay based on metal-enhanced luminescence and nanoparticle labeling amplification
  • Double-enhanced chemiluminescence immunoassay based on metal-enhanced luminescence and nanoparticle labeling amplification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Double Antibody Sandwich Method and Comparative Experiment

[0044] 1. Preparation of sensitized gold nanoparticles

[0045] 1. Preparation of gold nanoparticles with carboxyl groups on the surface of 1.120nm:

[0046] A. Prepare 1 mL of 4% chloroauric acid solution with deionized water.

[0047] B. Add 0.5mL of the above chloroauric acid solution into 200mL deionized water, stir well and bring to a boil.

[0048] C. Rapidly inject 3ml of 1% sodium citrate aqueous solution into the boiling solution.

[0049] D. Condensate and reflux for 30 minutes. The color of the suspension will change from dark blue to red as the gold nanoparticles form.

[0050] E. Cool to room temperature.

[0051] F. The particle size of the prepared gold nanoparticles measured under a transmission electron microscope is 13nm. The mass of a single spherical gold nanoparticle was calculated according to the particle size and density, and the particle concentration in the nano-gold ...

Embodiment 2

[0090] Embodiment 2: competition method and comparative experiment

[0091] 1. Use silicon dioxide to form an isolation layer on the surface of gold nanoparticles and couple E2 (estradiol) monoclonal antibody to the surface of the isolation layer: (the method here is the same as the NES sandwich method)

[0092] 1. Preparation of gold nanoparticles with 120nm surface hydroxyl groups:

[0093] A. Prepare 1 mL of 4% chloroauric acid solution with deionized water.

[0094] B. Add 0.5mL of the above chloroauric acid solution into 200mL deionized water, stir well and bring to a boil.

[0095] C. Rapidly inject 3ml of 1% sodium citrate aqueous solution into the boiling solution.

[0096] D. Condensate and reflux for 30 minutes. The color of the suspension will change from dark blue to red as the gold nanoparticles form.

[0097] E. Cool to room temperature.

[0098] F. The particle size of the prepared gold nanoparticles measured under a transmission electron microscope is 13nm. ...

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Abstract

The invention discloses a dual-enhanced chemiluminescent immunoassay method based on metal enhanced luminescence and nano particle labelled amplification. According to the dual-enhanced chemiluminescent immunoassay method based on metal enhanced luminescence and nano particle labelled amplification, metal nano particles replace conventional polystyrene nano microspheres, and luminescence substances are lebelled on the surface of an isolation layer spaced from the surfaces of the metal nano particles by 5-20nm; chemiluminescence of luminescence substances on sensitized metal nano particles is coupled to plasma waves on the surfaces of the metal nano particles; after resonance is generated, the chemiluminescence is emitted in the form of relatively high light intensity and relatively high attenuation speed. On the basis of a conventional chemiluminescence immunoassay technology, a metal enhanced luminescence technology and a nano particle labelled amplification technology are organically integrated; therefore, the dual-enhanced chemiluminescent immunoassay method has the advantages of high sensitivity, high detection speed and the like.

Description

technical field [0001] The invention belongs to the field of novel nanometer material research and medical examination for prevention and diagnosis, and relates to a double-enhanced chemiluminescence immunoassay method based on metal-enhanced luminescence and nanoparticle label amplification. Background technique [0002] Chemiluminescence immunoassay (CLIA), which combines high-sensitivity chemiluminescence assay technology with high-specificity immune response, is used for various antigens, haptens, antibodies, hormones, enzymes, fatty acids, vitamins and drugs and other detection and analysis techniques. It is a newest immunoassay technique developed after radioimmunoassay, enzyme immunoassay, fluorescent immunoassay and time-resolved fluorescent immunoassay. Compared with the traditional enzyme immunoassay, CLIA has the characteristics of higher sensitivity, shorter detection time, simpler labeling method and lower cost of raw materials. Nevertheless, with the gradual ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N21/76
CPCG01N21/76G01N33/54326
Inventor 金晶赵欢陈赢罗雅赛颜彬苏恩本
Owner GETEIN BIOTECH
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