Plastic-packaged colloidal gold detection card for detecting salmonella toxin

A Salmonella, colloidal gold technology, applied in the biological field, can solve the problems of complicated operation, long detection time, difficult to adapt to diagnosis and treatment, etc., and achieve the effect of convenient storage and prolonging the shelf life

Pending Publication Date: 2020-03-31
YANGZHOU UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Traditionally, the pathogen detection and identification methods of foodborne diseases in my country still remain in the traditional pathogen culture. The biggest weakness of this test method is that it is slow and difficult to adapt to diagnosis and treatment. It mainly includes two methods: one is PCR molecular identification technology to detect pathogens , although the sensitivity is high, fast, and can be detected directly, but the requirements for the laboratory and reagents are relatively high, otherwise the re

Method used

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  • Plastic-packaged colloidal gold detection card for detecting salmonella toxin
  • Plastic-packaged colloidal gold detection card for detecting salmonella toxin
  • Plastic-packaged colloidal gold detection card for detecting salmonella toxin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] 1) Salmonella toxin antigen purification

[0037] (1) Lysis and desalination of Salmonella toxin culture solution.

[0038] (2) Weak anion exchange chromatography: start buffer: 20mmol / L PBS, adjust the pH value to 6.0 with HCl. Elution buffer: 20mmol / L PBS, 1mol / L NaCl, HCl to adjust the pH value to 6.0. Equilibration: Connect Hiprep.16 / 10 DEAE FF weak anion exchange chromatography column to AKTA Explorer, rinse 100ml with elution buffer, and then equilibrate 100ml with start buffer. Flow rate 5ml / min. UV 280nm, 254nm, 215nm are monitored at the same time until the three curves are horizontal. Sample loading: Inject the desalted product into a 50ml sample loop at a flow rate of 5ml / min. Equilibrium: After the effluent peak is exhausted, use the start buffer to balance until the monitoring curve is horizontal. Elution: Salt gradient elution was used, and the eluent was elution buffer. The salt gradient is to make the concentration of NaCl rise from 0mol / L to 1mol / ...

Embodiment 2

[0053] A colloidal gold plastic-sealed detection card for detecting Salmonella toxin, the test strip includes a back plate, a sample pad, a colloidal gold pad, a nitrocellulose membrane and an absorbent pad, the sample pad, a colloidal gold pad, a nitrocellulose membrane and The absorbent pads are fixed on the backboard in sequence ( figure 1 ), the colloidal gold pad contains a colloidal gold-labeled anti-Salmonella toxin antigen monoclonal antibody; the nitrocellulose membrane is provided with a detection line and a quality control line. The test strip is sealed by cold lamination, the sample hole is located directly above the sample pad, and the sample loading area is exposed by tearing off the cover film or tearing the plastic seal ( figure 2 ). Direct observation after detection ( image 3 ).

Embodiment 3

[0055] Using the above method, a batch of products were trial-manufactured and sample tests were carried out.

[0056] A total of 50 samples of soil samples, sewage samples, and chicken manure samples were collected from the farm. Take 1 gram of sample respectively, add 2ml of deionized water and stir well, absorb the supernatant after sedimentation for 5 minutes, add 5 drops to 4ml of deionized water as the solution to be tested. Take the solution to be tested and add 4 drops to the sample pad of the test card, react for about 5 minutes and observe the results. As a result, 9 positive samples (18%) were detected, and the coincidence rate with the national standard method was 100%.

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a plastic-packaged colloidal gold detection card for detecting salmonella toxin and a preparation method of the plastic-packaged colloidal gold detection card. The detection card comprises a test strip, the test strip comprises a back plate, a sample pad, a colloidal gold pad, a nitrocellulose membrane and a waterabsorption pad, the sample pad, the colloidal gold pad, the nitrocellulose membrane and the water absorption pad are sequentially fixed on the back plate, and the colloidal gold pad contains a monoclonal antibody of a colloidal gold labeled salmonella toxin antigen; a detection line and a quality control line are arranged on the nitrocellulose membrane. Through the implementation of the process,a rapid production and preparation technology of the salmonella toxin rapid immunodetection card is developed.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a plastic-encapsulated colloidal gold detection card for detecting Salmonella toxin and a preparation method thereof. Background technique [0002] According to WHO estimates, the under-reporting rate of foodborne diseases in developed countries is more than 90%, while in developing countries it is more than 95%. Based on this inference, the food poisoning data currently available in my country is only the "tip of the iceberg" of foodborne diseases that actually occur in our country. For such a high rate of false positives, in addition to management problems, the limitation of detection and traceability methods of pathogenic microorganisms is also an important factor. At present, bacterial food poisoning mainly includes two types: infectious poisoning and toxin poisoning. Salmonella and its toxins are a common foodborne pathogen. [0003] At present, my country's food ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/569G01N33/558G01N33/532C07K16/12
CPCG01N33/577G01N33/56916G01N33/558G01N33/532C07K16/1235G01N2333/255G01N2469/10
Inventor 冀德君
Owner YANGZHOU UNIV
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