Colloidal gold detection test paper for ricin, preparation method thereof and application thereof

A technology for detecting ricin and test strips, which is applied in biological testing, measuring devices, material inspection products, etc., can solve the problems that cannot meet the needs of anti-terrorism, and achieve the effect of saving manpower and material resources

Inactive Publication Date: 2009-10-21
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This technology allows for quick detection by measuring tiny amounts of bacteria called rbc that are found inside food products or other materials being tested without having to manually count them yourself every time they go through production lines.

Problems solved by technology

This patented technical problem addressed in this patents relates to finding ways to quickly and accurately determine if or how much ricin was present at any given location without having access to expensive equipment such as X ray fluorescence analysis.

Method used

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  • Colloidal gold detection test paper for ricin, preparation method thereof and application thereof
  • Colloidal gold detection test paper for ricin, preparation method thereof and application thereof
  • Colloidal gold detection test paper for ricin, preparation method thereof and application thereof

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Effect test

Embodiment 1

[0051] Embodiment 1, the preparation of the colloidal gold test paper that detects ricin

[0052] 1. Preparation of colloidal gold probes

[0053] (1) HAuCl 4 It is first prepared as a 0.01% aqueous solution, and 100ml of pure water is taken and heated to boiling. Accurately add 1ml of 1% HAuCl under magnetic stirring 4 , while adding 1.5ml mass fraction of 1% trisodium citrate aqueous solution. After the color is stable, continue to heat and boil for 15 minutes. After cooling to room temperature, add pure water to make up to 100ml, and store in the dark at 4°C.

[0054] (2)K 2 CO 3 Adjust the PH value to about 8.0-8.5.

[0055] (3) Adjust the colloidal gold to an appropriate pH, preferably 8.0-8.5, more preferably 8.2, and dilute the antibody to 0.2 mg / ml with 5 mM PBS (pH 7.2).

[0056] 2. Preparation of gold-labeled antibody protective film

[0057] Take 1ml / tube of colloidal gold with pH 8.2 and distribute it in 1.5ml centrifuge tubes. Add the optimal labeling dos...

Embodiment 2

[0061] Embodiment 2, ricin detection test paper

[0062] Referring to Fig. 1, the reaction support is a 6.2cm × 0.4cm PCV plate; the absorbent pad is a 2cm × 0.4cm oil filter paper; the nitrocellulose membrane of 1.8cm × 0.4cm is coated with goat anti-mouse IgG (purchased from Dingguo Biotechnology Co., Ltd. company, 1mg / ml), rabbit anti-recombinant ricin A chain polyclonal antibody 1mg / ml (containing 1.5% BSA blocking agent), containing 0.4cm×0.4cm colloidal gold-labeled mouse anti-recombinant ricin A chain polyclonal Antibody (gold concentration: 12ug / ml diluted in 5mM PB; optimal pH value is 8.2) glass fiber membrane; the sample pad is a glass fiber membrane of 2.7cm × 0.4cm; that is, a ricin detection test paper is formed.

Embodiment 3

[0063] Embodiment 3, detection method

[0064] See attached figure 2 , mix the sample to be tested with the sample diluent, and then use a pipette to add the sample mixture to the sample hole of the test paper. The liquid in the sample goes up by siphon action, and the result is interpreted in 10-15 minutes:

[0065] If it contains ricin, it will form a corresponding complex with the colloidal gold-labeled mouse anti-recombinant ricin A chain polyclonal antibody on the test paper, and the upstream rabbit anti-recombinant ricin A chain polyclonal antibody coated on the nitrocellulose membrane Anti-binding, forming a red line, i.e. a red band at the "T".

[0066] Regardless of whether it contains the corresponding toxin or not, the colloidal gold-labeled polyclonal antibody continues to flow upward and forms a red precipitation line with the goat anti-mouse IgG coated on the membrane, that is, a red band is formed at "C". This line is a quality control line. If the colloidal g...

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Abstract

The invention discloses a detection test paper. The test paper comprises (1) a reaction support, (2) a water absorption pad, (3) a nitrocellulose membrane, (4) a gold-labeled antibody protective membrane and (5) a sample pad, wherein the nitrocellulose membrane is coated with a detection strip and a quality control strip of a rabbit anti-recombinant ricin A-link antibody and a quality control antibody; and the gold-labeled antibody protective membrane contains the rabbit anti-recombinant ricin A-link antibody labeled by colloidal gold. The test paper can be matched with a small instrument to carry out semi-quantitative detection, does not need professional training, has clear and identifiable result, can objectively store data, is simple to operate, easy to popularize, suitable for a basic level, suitable for field detection of emergency and suitable for epidemiological investigation, and plays an auxiliary role in diagnosis of ricin infection.

Description

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Claims

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Application Information

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Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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