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Immunochromatographic assay test paper for detecting staphylococcal enterotoxin B and preparation method thereof

The technology of immunochromatographic test paper and staphylococcus entero, which is applied in the directions of measuring devices, analytical materials, instruments, etc., can solve the problems such as the detection of B-type Staphylococcus aureus enterotoxin by immunochromatographic test paper, which is easy to handle. Effect

Inactive Publication Date: 2006-12-20
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0015] There is no report on the detection of type B Staphylococcus aureus enterotoxin by immunochromatographic test paper

Method used

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  • Immunochromatographic assay test paper for detecting staphylococcal enterotoxin B and preparation method thereof
  • Immunochromatographic assay test paper for detecting staphylococcal enterotoxin B and preparation method thereof
  • Immunochromatographic assay test paper for detecting staphylococcal enterotoxin B and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1, the preparation of the immunochromatography test paper that detects type B Staphylococcus aureus enterotoxin

[0043] 1. Preparation of specific antibody against Staphylococcus aureus type B enterotoxin

[0044] 1) Preparation of type B Staphylococcus aureus enterotoxin

[0045] Staphylococcus aureus (Staphylococcus aureus) FRI 243 was used for toxin-producing culture. The SEB strain was activated overnight at 37°C in the nutrient broth, and expanded the next day with 1% inoculum, and cultured at 37°C for 48 hours. Centrifuge at 8000 rpm at 4°C for 15 minutes, and collect the supernatant. The supernatant contains SEB, and the toxin production is about 100-200 μg / ml culture medium. 500ml of supernatant was diluted 10 times with 5mM PB pH7.0 buffer solution and added to SP-Sepharose HP column (the column was pre-equilibrated to pH7.0), and the sample loading flow rate was 5ml / min. Breakthrough peak UV2800mAU, equilibrate to UV60mAU with 5mM PB pH7.0 buff...

Embodiment 2

[0064] Embodiment 2, the detection of Staphylococcus aureus type B enterotoxin and cross-test with other related toxins

[0065] 1. Sensitivity detection of type B Staphylococcus aureus enterotoxin

[0066]1) SEB (1 mg / bottle) was prepared by the Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, and was used as a sample test solution after limiting dilution with normal saline.

[0067] 2) Detected by the immunochromatographic test paper coated with SEB-specific antibody and goat anti-rabbit IgG prepared in Example 1, add 3 drops of sample detection solution (about 150 μl), start to observe the results after 2 minutes, and stop the observation after 15 minutes. Detect different concentrations of SEB, above 10ng / ml can be detected (see Table 3), compared with other immunological methods have higher sensitivity.

[0068] SEB sample concentration

5ng / ml

10ng / ml

20ng / ml

40ng / ml

80ng / ml

160ng / ml

Test resu...

Embodiment 3

[0076] Embodiment 3, SEB rapid detection reagent is to the detection of SEB in the simulated environment specimen

[0077] 1. Preparation of mock specimens

[0078] Select ham sausage, milk, human serum, ordinary broth medium and soil simulation environment field samples, weigh 2g of ham sausage respectively, cut into pieces and put them in a container, add 5ml of sterile normal saline, stir well, let stand for precipitation; Milk is diluted with normal saline 1:4; serum is diluted with normal saline 1:10; stock solution is used for ordinary broth; 1g of soil is weighed, added with 5ml of sterile normal saline, and centrifuged at 2000rpm for 10-15 minutes.

[0079] Each sample was made in multiple copies, and the supernatant of one copy was used as a negative control, and the other samples were added with a known concentration of pure SEB, so that the samples contained SEB 20ng / ml, 40ng / ml, 80ng / ml and 1600ng / ml respectively, as a simulation positive test specimen.

[0080] ...

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Abstract

The invention discloses an immune chromatographic indicator paper and making method of B-typed staphylococcus aureus enterotoxin, which comprises the following parts: sample pad 1, metal pad 2 with specific antibody mark colloidal gold probe of B-typed staphylococcus aureus enterotoxin in connection with one end of sample pad tightly, nitric fiber film (NC film) 3 in connection with the other end of metal pad tightly, water-absorbing pad 4 in connection with the other end of nitric fiber film tightly, wherein the nitric fiber film covers mutually separated detecting line 5 and quality control line 6, which is specific antibody of B-typed staphylococcus aureus enterotoxin and sheep-anti-rabbit lgG separately.

Description

technical field [0001] The invention relates to a test paper for detecting type B Staphylococcus aureus enterotoxin and a preparation method thereof, in particular to an immunochromatographic test paper for detecting type B Staphylococcus aureus enterotoxin, and a preparation method thereof. Background technique [0002] Staphylococcal enterotoxin (SE) is a protein exotoxin produced by Staphylococcus aureus and can be divided into multiple serotypes A to I. Among them, Staphylococcus enterotoxin B (SEB) has the highest toxin production (Gao Shude, Wang Meixian, editor-in-chief, Handbook of Epidemic Prevention and Inspection, p83-85, 1982, People's Medical Publishing House). Since SEB is an important incapacitating toxin, it is included in the verification list of the International Convention for the Prohibition of Biological Weapons as a classic biological warfare agent. After the "9.11 Incident", the United States listed SEB, along with Bacillus anthracis and other potent ...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/558G01N33/532
Inventor 端青姜永强郑裕玲檀华朱虹何君
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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