Visualized enzyme linked immunoassay method

An enzyme-linked immunoassay and immunoassay technology, applied in the field of visual enzyme-linked immunoassay, can solve the problems of low detection limit, fast reaction time, and single color change, and achieve low detection limit, fast reaction time, and color development. Sensitive effect

Inactive Publication Date: 2017-12-19
FUZHOU UNIV
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Problems solved by technology

The visual immunoassay method of the present invention can produce multiple color changes, solves the shortcoming of single color change in the traditional colorimetric method, can realize the quantitative detection of the target object, and h...

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  • Visualized enzyme linked immunoassay method
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  • Visualized enzyme linked immunoassay method

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Embodiment 1

[0025] The following embodiments illustrate the operation process of applying the present invention to carry out actual sample analysis in conjunction with the accompanying drawings:

[0026] (1) Before the synthesis, soak all the glassware needed in aqua regia, then rinse with a large amount of water, and finally rinse with ultrapure water;

[0027] (2) Synthesis of gold nanobicones with LSPR peak of 800nm: 0.25 mL of 25 mM NaBH 4 The solution was added to 10 mL of a mixed solution containing 0.25 mM HAuCl to form a brown-yellow solution after 2 minutes 4 , 50 mM CTAC and 5 mM citric acid; the resulting brown-yellow solution was placed in an oil bath at 80°C for 90 minutes to obtain a gold seed solution, which was stored at room temperature for subsequent use;

[0028] Then prepare the growth solution, that is, put 10 mL 0.01 M HAuCl 4 solution, 2 mL 0.01 M AgNO 3solution, 4 mL of 1M HCl solution and 1.6 mL of 0.1 M AA solution were added to 200 mL of 0.1 M CTAB solution. ...

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Abstract

The invention discloses a visualized enzyme linked immunoassay method which comprises the following steps: (1) Au NBP@Ag nanorod preparation; (2) enzyme-linked immunoassay. According to the visualized enzyme linked immunoassay method, a primary antibody has been laid on a plate, antigen is added, then catalase modified secondary antibody compound is added, and then Fe2<+> and hydrochloric acid are added after the equivalent of hydrogen peroxide is added; then a silver plated gold cone nanorod is added to react, a series of fresh colors are observed in an etching process, and a target antigen is analyzed visually and semi quantitatively. The visualized enzyme linked immunoassay method disclosed by the invention can generate varieties of color change, solves the defect that color change in a traditional colorimetric method is single, achieves quantitative detection on a target object and further has quick reaction time and a good effect; thus, accuracy of the method for naked eye detection is greatly improved, and a detection limit is lower.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, and in particular relates to a visual enzyme-linked immunoassay method. Background technique [0002] In recent years, great attention has been paid to various colorimetric sensors due to their unique advantages in terms of simplicity. Usually, colorimetric sensors can be detected with ordinary UV-vis spectrometers, and can also be observed with naked eyes. Due to their simplicity, these sensors are used to detect various biological and biomedical targets. The most commonly used colorimetric reactions are horseradish peroxidase (HRP) and 3,3′,5,5′-tetramethylbenzidine (TMB) immunosystems, which have been used to detect many disease markers Based on the specific recognition of antibody-antigen and the efficient color reaction between HRP and TMB. In addition to TMB, some other chromogenic substrates, such as o-phenylenediamine (OPD) and 2'-hydrazine-bis-3-ethylbenzothiazoline-6-sulfonic acid...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/553
CPCG01N33/54346G01N33/553
Inventor 郭隆华林艺林振宇邱彬陈国南
Owner FUZHOU UNIV
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