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Colloidal gold test paper for staphylococcal enterotoxin B (SEB), preparation method thereof and application thereof

A technology of Staphylococcus aureus enterotoxin and colloidal gold, applied in biological testing, material inspection products, measuring devices, etc., can solve the problems of long detection time, cumbersome operation, unsuitable for on-site detection, etc., 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

Problems solved by technology

At present, there is an ELISA method for the determination of Staphylococcus aureus enterotoxin type B, which needs to be completed in the laboratory with the aid of a Meibiao instrument. [J] Journal of Cellular and Molecular Immunology, 1007-8738 (2007) 08-0761-02); animal toxicology experiments are cumbersome to operate, and the detection time is long, requiring rich experience and designated experimental environment. Suitable for on-site testing

Method used

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  • Colloidal gold test paper for staphylococcal enterotoxin B (SEB), preparation method thereof and application thereof
  • Colloidal gold test paper for staphylococcal enterotoxin B (SEB), preparation method thereof and application thereof
  • Colloidal gold test paper for staphylococcal enterotoxin B (SEB), preparation method thereof and application thereof

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Experimental program
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Effect test

Embodiment 1

[0044] Embodiment 1, the preparation of the colloidal gold test paper that detects Staphylococcus aureus enterotoxin B type

[0045] 1. Preparation of colloidal gold probes

[0046] (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.

[0047] (2)K 2 CO 3 Adjust the PH value to about 8.2-8.5.

[0048] (3) Adjust the colloidal gold to a suitable pH, preferably 8.0-8.5, more preferably 8.2, and dilute the antibody to 0.2 mg / ml with 5 mM PB (pH 7.2).

[0049] 2. Preparation of gold-labeled antibody adsorption membrane

[0050] Take 1ml / tube of colloidal gold with pH 8.2 and distribute it in 1.5ml centrifuge tu...

Embodiment 2

[0054] Embodiment 2, Staphylococcus aureus enterotoxin type B detection test paper

[0055] Referring to Figure 1, the reaction support is a 6.2cm × 0.4cm PCV plate; the absorbent pad is 2cm × 0.4cm oil filter paper; the nitrocellulose membrane of 1.8cm × 0.4cm is coated with goat anti-rabbit IgG (purchased from Dingguo Biotechnology company, 1mg / ml), rabbit anti-SEB polyclonal antibody 2mg / ml (containing 1% BSA blocking agent), containing 0.4cm×0.4cm colloidal gold-labeled rabbit anti-SEB polyclonal antibody (concentration of standard gold: 5ug PBS dilution; Optimum pH value is 8.5) glass fiber membrane; sample pad is the glass fiber membrane of 2.7cm * 0.4cm; That is to form the staphylococcus aureus enterotoxin type B detection test paper.

Embodiment 3

[0056] Embodiment 3, detection method

[0057] 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:

[0058] If it contains Staphylococcus aureus enterotoxin type B antigen, it will form a corresponding complex with the colloidal gold-labeled rabbit anti-SEB polyclonal antibody on the test paper, and combine with the flow rabbit anti-SEB polyclonal antibody coated on the nitrocellulose membrane to form a red color. line, that is, a red band is formed at the "T".

[0059] Regardless of whether it contains the corresponding antigen or not, the colloidal gold-labeled polyclonal antibody continues to crawl upward and forms a red precipitation line with the goat anti-rabbit IgG coated on the membrane, that is, a red band is formed at "C". This line is a quality control line...

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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 adsorption membrane and (5) a sample pad, wherein the nitrocellulose membrane is coated with a detection strip and a quality control strip of a staphylococcal enterotoxin B antibody and a quality control antibody; and the gold-labeled antibody adsorption membrane contains the staphylococcal enterotoxin B 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 staphylococcal enterotoxin B infection.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a test paper for detecting staphylococcus aureus enterotoxin type B (SEB), a preparation method thereof and an application in staphylococcus aureus enterotoxin type B (SEB). Background technique [0002] Staphylococcus aureus (Staphylococcus aureus) is the main pathogenic bacterium that causes infection and toxic diseases, and its pathogenicity is mainly related to the production of various pathogenic substances, Staphylococcal en2terotoxin B (SEB) is one of them An important pathogenic substance. At present, there is an ELISA method for the determination of Staphylococcus aureus enterotoxin type B, which needs to be completed in the laboratory with the aid of a Meibiao instrument. [J] Journal of Cellular and Molecular Immunology, 1007-8738 (2007) 08-0761-02); animal toxicology experiments are cumbersome to operate, and the detection time is long, requiring rich e...

Claims

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Application Information

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IPC IPC(8): G01N33/551G01N33/552G01N33/532G01N33/558G01N33/52
Inventor 王静姜永强杨宇张晓龙孙肖红胡孔新
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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