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Colloidal gold immuno-chromatographic assay for quantitatively detecting staphylococcal enterotixn B and gold-immuochromatography assay test paper

A technology for the detection of staphylococcus aureus enterotoxin and test paper, which is applied in the direction of measuring devices, analysis materials, instruments, etc., and can solve the problem of fewer products detected in the field of food hygiene

Inactive Publication Date: 2010-03-10
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, this method has developed rapidly and has been widely used in the field of biomedicine, especially in medical testing, but there are few products used in the field of food hygiene testing.

Method used

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  • Colloidal gold immuno-chromatographic assay for quantitatively detecting staphylococcal enterotixn B and gold-immuochromatography assay test paper
  • Colloidal gold immuno-chromatographic assay for quantitatively detecting staphylococcal enterotixn B and gold-immuochromatography assay test paper
  • Colloidal gold immuno-chromatographic assay for quantitatively detecting staphylococcal enterotixn B and gold-immuochromatography assay test paper

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1: the preparation of colloidal gold immunochromatography test strip

[0035]1. Antigen and antibody

[0036] Staphylococcus aureus enterotoxin B and its antibody (SEB recombinant antigen, rabbit polyclonal antibody, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences), goat anti-rabbit IgG (purchased from Dingguo Biotechnology Company)

[0037] 2. Consumables for chromatography test strips

[0038] Bonding pad (glass fiber), nitrocellulose membrane (NC membrane, SHF 1350225), sample pad, absorbent pad, and filter paper were purchased from Minipore.

[0039] 3. Experimental equipment

[0040] Gold Standard Immunity Analyzer (jointly developed by Shanghai Institute of Optics and Fine Mechanics, Chinese Academy of Sciences, and China Academy of Inspection and Quarantine).

[0041] 4. Preparation of colloidal gold immunochromatographic test strips

[0042] 4.1 Colloidal gold bonding pad

[0043] Label the rabbit anti-SEB antibod...

Embodiment 2

[0048] Embodiment 2: Qualitative and quantitative detection of samples

[0049] 1. Sample processing

[0050] Use the sample treatment solution (5mM PBS pH7.4) to dilute animal serum, milk and other viscous liquid samples 1:20, add solid food such as ham sausage at 1 / 100 (W / V) to the sample treatment solution to dissolve, stand still and take the Clear, stand still and take the supernatant. Different doses of natural Staphylococcus aureus enterotoxin B (SEB) were added as mock test samples.

[0051] 2. Qualitative detection

[0052] Add 100ul of the treated sample and sample treatment solution (as a negative sample) to the end of the prepared chromatographic strip sample pad, let it stand for 15min, and observe the result. If both the test strip and the quality control strip appear red, it is judged as positive, and if only the quality control strip appears red, it is negative.

[0053] 3. Quantitative detection

[0054] 3.1. Determination of judgment value

[0055] Acco...

Embodiment 3

[0058] Embodiment 3: sensitivity test

[0059] 1. Sensitivity of quantitative detection

[0060] Detect different concentrations of natural Staphylococcus aureus enterotoxin B (SEB), the calculated judgment value (CUT-OFF) is 0.0243, the average The values ​​of the T / C ratio are all 0.00, lower than 0.043, which is negative; the average T / C ratios of 10ng / ml~10ug / ml natural Staphylococcus aureus enterotoxin B (SEB) are higher than The result of 0.0243 is positive; when the concentration of natural Staphylococcus aureus enterotoxin B (SEB) is greater than 10ng / ml, the average value of T / C ratio is greater than 0.0243, so the detection sensitivity is 10ng / ml.

[0061] Table 1 Detection results of various concentrations of Staphylococcus aureus enterotoxin B (SEB) colloidal gold immunochromatography test strips

[0062] serial number

Detection value T

(V)

Quality control value C

(V)

T / C

Interpretation of results

0

0.0...

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Abstract

The invention provides a colloidal gold immuno-chromatographic assay for detecting staphylococcal enterotixn B (SEB). The colloidal gold immuno-chromatographic assay can rapidly, sensitively, specifically and accurately detect the staphylococcal enterotixn B (SEB) in a sample and realize quantitative detection, and is suitable for on-spot rapid detection.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a rapid quantitative detection method for the detection of Staphylococcus aureus toxin B and a colloidal gold immunological detection test strip. 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 its production of various pathogenic substances, Staphylococcal en2terotoxin B (SEB) is one of them Important pathogenic substance, its mouse LD value is 0.023-5μg / kg. Staphylococcus aureus can cause food poisoning, the main reason is that it can produce enterotoxin (Enterotoxin, referred to as SE). About 1 / 3 of the Staphylococcus aureus isolated from the clinic produce enterotoxin. Staphylococcus aureus enterotoxin is mainly a kind of extracellular protein with related structure, similar virulence and different antigenicity produ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/558G01N33/552G01N33/532
Inventor 王静姜永强张晓龙杨宇孙肖红胡孔新姚李四
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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