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Fast chemiluminescence immune detection system and analysis method

A chemiluminescent immunoassay and detection system technology, which is applied in the field of double acceleration of the incubation process of immunoassay by infrared heating and turbulent mixing, and in the field of automatic detection instrument system, can solve the problem of slow heating speed, increased cost, limited fast and low-cost immunoassay Improve the reproducibility, reduce the manual operation, and reduce the cost of analysis

Inactive Publication Date: 2014-05-14
NANJING UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods have the following disadvantages: (1) Usually, the internal diameter of the pipeline of the flow injection analysis system is about 1 mm, so at low flow rates, the fluid in the pipeline is laminar flow
However, due to the slow temperature rise of water bath heating and the need to configure a water bath magnetic stirrer, the cost is increased, which limits the development of fast and low-cost immunoassay methods to a certain extent.

Method used

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  • Fast chemiluminescence immune detection system and analysis method
  • Fast chemiluminescence immune detection system and analysis method

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Embodiment 1 is combined with attached figure 1 Further explanation of the chemiluminescent detection system:

[0034] The system consists of a solution transmission system, an incubation system, a magnetic separation system, and a signal acquisition system. Wherein the solution transmission system consists of the first small test tube (1), the second small test tube (2), the third small test tube (3), the fourth small test tube (4), multi-position valve (5), peristaltic pump (6) and Silicone rubber tube (7) is formed. The first small test tube (1) is equipped with the sample to be tested, the enzyme-labeled antibody and the magnetic bead mixture with the capture antibody immobilized on the surface, and the second small test tube (2) is equipped with 0.01M phosphate washing buffer (pH7.4, containing 0.05% Tween-20), the third small test tube (3) is equipped with chemiluminescent substrate solution (luminol, hydrogen peroxide and p-iodophenol), and the fourth small tes...

Embodiment 2

[0036] Example 2 Detection steps of the chemiluminescence immunoassay system

[0037] The specific analysis process is shown in Table 1.

[0038]

[0039] All analysis steps are programmed and automatically controlled by computer, and the measured chemiluminescent signal is output from the computer. After the analysis steps were completed, the magnetic beads immobilized with antibodies were collected uniformly, soaked in glycine / hydrochloric acid buffer solution with pH 2.2 for 10 minutes, and washed with 0.01M phosphate buffer solution (pH 7.4) for reuse.

Embodiment 3

[0041] Taking alpha-fetoprotein (AFP), a marker with significant significance in the clinical diagnosis of tumors, as an example, the application of this automatic sample injection resolution chemiluminescence multicomponent immunoassay method is illustrated.

[0042] Using epoxy-activated magnetic beads as the carrier of the immune reaction, the mouse monoclonal AFP antibody was covalently immobilized, and the residual active sites were blocked with bovine serum albumin. The tracer antibody was goat polyclonal AFP antibody labeled with horseradish peroxidase.

[0043] As shown in the process shown in Table 1, the tumor marker AFP in clinical serum samples was used as the detection object, and the chemiluminescence immunoassay was performed to obtain the concentration of AFP in the test sample, which provided a feasible method for clinical screening and diagnosis.

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Abstract

The invention relates to a fast chemiluminescence immune detection system and an analysis method. The solution transmission system of the detection system consists of cuvettes (1-4), a multi-position valve (5), a peristaltic pump (6) and a silicone rubber tube (7); an incubation system consists of a glass helix tube (8) and a vacuum infrared heating tube (9); a magnetic separation system consists of a glass coiled tube (10) and a permanent magnet (11); and a signal collection system consists of a glass coiled tube (12) and a photomultiplier (13). All the systems are connected by polytetrafluoroethylene tubes. Based on the sandwich method immune analysis and the flow injection system of magnetic bead surfaces, the invention carries out the infrared heating for the incubation process of the immune analysis, i.e. a turbulent mixing double acceleration method, so that the whole analysis process is reduced to 3 minutes and the inspection process can be controlled automatically. The method has the characteristics of fastness, simplicity, convenience, good reproducibility, high sensitivity, low cost and the like, can be applied to fields such as clinical diagnosis, environment monitoring, food safety and the like.

Description

1. Technical field [0001] The invention is a chemiluminescence immunoassay technique, which relates to the dual acceleration of the incubation process of the immunoassay by infrared heating and turbulent mixing. Using magnetic beads as immunoreaction carrier, based on surface sandwich immunoassay and flow injection chemiluminescence analysis technology. The invention also relates to an automatic detection instrument system for chemiluminescence immunoassay. 2. Background technology [0002] Immunoassay has the advantages of high selectivity, high sensitivity, and good universality, and has a wide range of applications in the fields of food safety, environmental monitoring, clinical diagnosis, drug analysis, and microbial testing. When immunology is applied to practical fields, rapid, low-cost and automated screening of a large number of samples is often required. For example, early diagnosis of diseases, food safety inspection and environmental monitoring all require fast,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/53G01N21/76G01N33/551
Inventor 刘宏严枫鞠熀先
Owner NANJING UNIV
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