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

A technology for detection of acacia toxin and test paper, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of not being able to meet the needs of anti-terrorism, difficulty in obtaining antibodies to highly toxic toxins, and long incubation time of ELISA method, so as to save manpower Material effect

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

AI Technical Summary

Problems solved by technology

[0005] Although the above method has high sensitivity, it has great limitations for the detection of samples: first, it is difficult to obtain antibodies to highly toxic toxins, and radioimmunoassay also requires at least part of the primary structure of the analyte to be known; in addition, the incubation time of the ELISA method Long, not suitable for on-site testing
The above methods can no longer meet the needs of contemporary anti-terrorism, and there is an urgent need for on-site, rapid and quantitative detection methods of abrin toxin

Method used

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

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Experimental program
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Effect test

Embodiment 1

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

[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 standard pad

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

Embodiment 2

[0061] Embodiment 2, abrin toxin 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 abrin toxin A chain polyclonal antibody 1mg / ml (containing 1.5% BSA blocking agent), containing 0.4cm×0.4cm colloidal gold-labeled mouse anti-recombinant abrin toxin A chain polyclonal Antibody (standard gold concentration: 12ug / ml diluted in 5mM PBS; optimal pH value is 8.2) glass fiber membrane; the sample pad is a glass fiber membrane of 2.7cm×0.4cm; that is, an abrin toxin 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 abrin toxin, it will form a corresponding complex with colloidal gold-labeled mouse anti-recombinant abrin toxin A chain polyclonal antibody on the test paper, and go upstream with rabbit anti-recombinant abrin toxin A chain polyantibody coated on nitrocellulose membrane Combined, a red line is formed, 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 colloi...

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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 and (4) a gold-labeled antibody protective membrane, wherein the nitrocellulose membrane is coated with a detection strip and a quality control strip of a rabbit anti-recombinant abrin A-link antibody and a quality control antibody; and the gold-labeled antibody protective membrane contains the rabbit anti-recombinant abrin 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 abrin infection.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a test paper for detecting abrin toxin, a preparation method thereof and an application in detecting abrin toxin. Background technique [0002] Abrin toxin is a highly toxic high-molecular protein toxin extracted from the seeds of the leguminous vine plant Abrusprecatorius, and its content accounts for about 2.8% to 3.0% of the seeds. There are four homologous toxins (isoabrins) of abrin toxin, namely abrin-a, abrin-b, abrin-c, and abrin-d, with relative molecular masses between 63kDa and 67kDa, which are encoded by different genes, but Both belong to a multi-gene family; among them, abrin-b and abrin-c have only weak cytotoxicity due to their low B-chain agglutinating activity; while abrin-a and abrin-d have extremely strong cytotoxicity. LD50 is 0.04μg / kg, the lethal dose ingested by adults is 5.0μg / kg~7.0μg / kg, its toxicity intensity is more than 70 times that ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/552G01N33/532G01N33/558
Inventor 王静王景林聂聪杨宇孙肖红高姗胡孔新张晓龙
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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