A kind of high-sensitivity colloidal gold test paper and its preparation method and application

A colloidal gold test paper, a high-sensitivity technology, applied in the biological field, can solve the problems of markers and gold particles falling off, compound instability, etc., and achieve the effect of simple preparation method, good market prospect, and improved detection sensitivity

Active Publication Date: 2018-09-14
WUHAN J H BIO TECH
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  • Claims
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Problems solved by technology

However, this method may have a false positive problem in actual operation, because the preparation process of the streptavidin-gold-antibody conjugate is to directly add the antibody and avidin to the colloidal gold solution for labeling. There are certain technical loopholes in adding two substances to colloidal gold for labeling at the same time, because there are certain differences in the labeling conditions of different substances. The labeling method of this method will produce avidin-gold, antibody-gold and avidin-gold. Gold-antibody three substances, and because the same labeling conditions are used, the complex in the above-mentioned markers is unstable, and the markers and gold particles are prone to fall off during the storage or use of the test paper, resulting in There are free colloidal gold particles, and the free colloidal gold will directly react with the detection antibody on the NC membrane of the test strip during the detection, so there is a risk of false positive

Method used

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  • A kind of high-sensitivity colloidal gold test paper and its preparation method and application

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Embodiment 1

[0029] Preparation of High Sensitivity Colloidal Gold Test Paper

[0030] 1. Preparation of Colloidal Gold Solution

[0031] Accurately measure 1000mL of purified water, and quickly add 10mL of gold chloride solution after boiling; after the solution boils again, quickly add 10mL of colloidal gold reducing agent, continue to boil, and observe that the color of the solution changes from yellow to black and then to purple, and finally becomes After the stable wine red color, continue to boil for 10 minutes by timing, stop heating, and after the solution is cooled to room temperature, add purified water to make up to 1000mL.

[0032] 2. Antibody pretreatment before biotinylation

[0033] Cut the dialysis bag into small pieces of 10-20cm; in 500mL of 2% NaHCO 3 Boil the dialysis bag in 1mM EDTA·2Na (pH 8.0) for 10min; wash thoroughly with distilled water; place the dialysis bag in 500mL of 1mM EDTA·2Na (pH 8.0) and boil for 10min; after cooling, place the dialysis bag in distill...

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Abstract

The invention discloses a high-sensitivity colloidal gold test paper and a preparation method and application thereof. The high-sensitivity colloidal gold test paper comprises a base plate, a sample pad, a conjugate pad, an NC (Nitrocellulose) membrane and an absorbent pad, wherein the conjugate pad is a conjugate pad containing a gold standard biotin antibody; and the sample pad is a sample pad containing an avidin. The high-sensitivity colloidal gold test paper is applicable to detection on various antibodies / antigens and has high flexibility and low false positive rate. Compared with the traditional colloidal gold test paper, the high-sensitivity colloidal gold test paper has the advantages that the detection sensitivity is improved by 100 multiples, the preparation method is simple, the practicability is good, and the market prospect is relatively good.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a high-sensitivity colloidal gold test paper and its preparation method and application. Background technique [0002] Colloidal gold is a commonly used labeling technology. It is a new type of immunolabeling technology that uses colloidal gold as a tracer marker to apply to antigens and antibodies. It has its unique advantages. It has been widely used in various biological researches in recent years. Almost all immunoblotting techniques used clinically use its markers. At the same time, it may be used in flow cytometry, electron microscopy, immunology, molecular biology and even biochips. [0003] In order to improve the sensitivity of colloidal gold detection, researchers have adopted various methods, including the biotin-avidin system (Biotin-Avidin-System, BAS), which was developed in the late 1970s. Novel biological reaction amplification system. With the advent of various b...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/558
CPCG01N33/558
Inventor 周军王健斌龚贻洲曾生
Owner WUHAN J H BIO TECH
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