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Enzyme-linked immunosorbent assay sandwich method kit for detecting antigen of animal pestis F1

An enzyme-linked immunosorbent (ELISA) and F1 antigen technology, which is used in measurement devices, instruments, scientific instruments, etc., can solve the problems of wrong test results and inability to obtain positive results, and achieves the reduction of reaction steps, the improvement of specificity, and the safety of the detection process. Effect

Inactive Publication Date: 2010-09-22
THE CENT FOR DISEASE CONTROL & PREVENTION OF XINJIANG UYGUR AUTONOMOUS REGION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are professional organizations in my country to monitor the plague in the epidemic foci. The "National Plague Surveillance Program" monitors plague in animals using the isolation and culture of Y. pestis and the reverse-phase indirect hemagglutination test to detect the plague F1 antigen; the isolation and culture of fresh samples of Y. pestis takes 24 hours at the fastest The results can only be obtained, and the isolation and culture method can only detect live bacteria of Yersinia pestis, and cannot obtain positive results from animal specimens infected with plague that have been used with antibiotics, contaminated with fungicides, or corrupted old; F1 antigen is the specific antigenic component of Yersinia pestis, The reversed-phase indirect hemagglutination test detects the plague F1 antigen, non-specific agglutination often occurs in spoiled samples, and sometimes high-titer non-specific agglutination covers up positive reactions, resulting in wrong test results

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: Preparation of ELISA sandwich method kit for detecting animal plague antigen

[0028] 1. Preparation of ELISA sandwich method kit for detection of animal plague antigen

[0029] (1) The composition of the ELISA sandwich method kit for detecting animal plague antigens:

[0030] 1. Plague F1 antibody-coated plate, that is, F1Ab plate, 8×12 tube plate, 1 piece

[0031] 2. Horseradish peroxidase-labeled plague F1 antibody (HRP-F1Ab): freeze-dried 1 tube

[0032] 3. Glycerin. PB 2ml 1 stick

[0033] 4.10×PBS.T 20ml 1 bottle

[0034] 5. Plague F1 antigen positive control: 2ml 1 tube

[0035] 6. Negative control: 1 tube of 2ml

[0036] 7. Plague F1 antibody for inhibition: freeze-dried 1 tube

[0037] 8. TMB Chromogenic Solution: A liquid 6ml 1 bottle B liquid 6ml 1 bottle

[0038] 9. Chromogenic termination solution is 10% H 2 SO 4 : 6ml 1 bottle

[0039] 10.1% Thimerosal: 2ml 1 stick

[0040] (2) Preparation method of each component:

[0041] 1. P...

Embodiment 2

[0071] Embodiment 2: the application method of the ELISA sandwich method kit that detects animal plague antigen

[0072] 1. Preparation of reagents for detection of animal plague F1 antigen by ELISA sandwich method

[0073] 1. PBS.T: 10×PBS.T diluted 10 times with 0.85% NaCl.

[0074] 2. HRP-Fl antibody storage solution: add glycerol-PB2m1 to dissolve each HRP-Fl antibody (lyophilized agent), mix well, -20 0 C storage, shelf life of 6 months.

[0075] HRP-Fl antibody application solution: Dilute HRP-Fl antibody stock solution 30 times with PBS.T, 4 0 C kept for 10 days.

[0076] 3. F1 antigen positive control: Dissolve each F1 antigen positive control (lyophilized agent) with PBS.T2m1 and mix well.

[0077] 4. Negative control: Dissolve each negative control (lyophilized agent) in PBS.T2m1 and mix well.

[0078] 5. Plague F1 antibody for inhibition: dissolve and mix with PBS.T5m1.

[0079] 6. TMB Chromogenic Solution: Mix equal amounts of Chromogenic Solution A and B, an...

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PUM

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Abstract

The invention relates to the technical field of the epidemic surveillance and the diagnosis of animal pestis, in particular to an enzyme-linked immunosorbent assay sandwich method kit for detecting the antigen of animal pestis F1. The Kit comprises an enzyme labelled antibody, an antibody-coated plate, glycerol, 2mL of PB liquid, 20mL of 10*PBS.T, 2mL of positive control of antigen of the animal pestis F1, 2mL of negative control, 5mL of antibody of the pestis F1 for restraining, 6mL of TMB coloration liquid A liquid, 6mL of TMB coloration liquid B liquid, 6mL of coloration stopping liquid and 2mL of 1% thiomersalate. The kit has the beneficial effects of having a safe process since a material to be detected is the antigen of the pestis F1 which can be detected after being sterilized with bactericide, improving the specificity of the detection since the antigen to be detected of the pestis F1 is specifically and selectively combined with the coating antibody and the enzyme labelled antibody for twice, reducing the step of enzyme labeling a second antibody enzyme label, reducing the non-specific coloration, having the meaning of quantifying by detecting a result with an elisa meter, creating a convenient condition for a field site without a power source by visually inspecting a judgment result, increasing a procedure of a restraining test, and improving the accuracy of the detecting result.

Description

technical field [0001] The invention relates to the technical field of animal plague epidemic monitoring and plague diagnosis, and relates to an enzyme-linked immunosorbent assay sandwich method kit for detecting animal plague F1 antigen. Background technique [0002] Plague is one of the Class A infectious diseases stipulated in my country's "Law on the Prevention and Control of Infectious Diseases". It is a severe zoonotic infectious disease caused by Yersinia pestis. In my country, 12 types of natural plague foci have been found, distributed in 19 provinces, 292 counties and cities, covering an area of ​​1.31 million km 2 . Since the 1990s, animal plague has continued to be active, and human plague has occurred year after year. The risk of long-distance and high-speed transmission of plague has increased. Rodents are the main host animal and source of infection of plague, which can naturally infect nearly 200 species of plague animals. There are professional organizati...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/535G01N33/543
Inventor 热娜.吐尔地雷刚蒋卫闫化奎徐兵唐建国
Owner THE CENT FOR DISEASE CONTROL & PREVENTION OF XINJIANG UYGUR AUTONOMOUS REGION
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