A detection card for rapid detection of canine distemper virus antigen and its preparation method

A technology of canine distemper virus and detection card, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problem of inability to effectively distinguish antibodies and antibodies, and achieve the effect of simple, efficient and sensitive detection suitable for popularization and use.

Active Publication Date: 2022-06-21
SHANDONG LVDU BIO SICIENCE & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can realize rapid on-site detection of canine distemper virus antibody, but this method can only be used to detect the antibody content in serum, and cannot effectively distinguish whether it is the antibody produced by virus infection or the antibody produced after vaccination

Method used

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  • A detection card for rapid detection of canine distemper virus antigen and its preparation method
  • A detection card for rapid detection of canine distemper virus antigen and its preparation method
  • A detection card for rapid detection of canine distemper virus antigen and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: a kind of preparation method of rapid detection canine distemper virus pathogen detection card, concrete steps are:

[0036] (1) Isolation, culture and purification of canine distemper virus.

[0037] Three regions including Shandong, Guangdong, and Sichuan were selected and confirmed by PCR as strong positive eye secretions, inoculated into F81 cells for 3 generations blindly, and the cells with significant lesions were collected, cryopreserved, and expanded for culture, collection, and ultrafiltration concentration. 10 times, then repeated freeze-thaw 3 times, centrifuged at 10,000 rpm for 30 minutes, applied a sucrose density gradient, 4 gradients of 30%, 35%, 40%, 45%, and centrifuged at 26,000 rpm for 30 minutes. content.

[0038] (2) Preparation of anti-canine distemper virus paired monoclonal antibodies

[0039] ①Immunization program

[0040] The purified virus of about 1mg / ml from the three regions was used as the immunogen and 501 adjuvant was ...

Embodiment 2

[0060] Embodiment 2: Pretreatment method and detection method of tested sample

[0061] The using method of the canine distemper virus pathogen rapid detection card of the present invention is as follows:

[0062] a. Collect dog, mink, fox eye secretions or nasal fluid and saliva with cotton swabs.

[0063] b. Open the sample collection tube and insert a cotton swab.

[0064] c. Agitate the swab well until the sample on the swab is dissolved in the diluent, then discard the swab.

[0065] d. After shaking and mixing, vertical sample tube, aspirate the supernatant and drop 3-5 drops into the sample hole in the detection card.

[0066] e. The judgment result is judged at 5 minutes, and the judgment at other time is invalid.

[0067] (1) Positive ( figure 2 ): If the C line develops color and the T line develops color, it is judged as positive.

[0068] (2) negative ( image 3 ): If the C line develops color and the T line does not develop color, it is judged as negative....

Embodiment 3

[0070] Example 3: Test Card Performance Measurement

[0071] (1) Stability

[0072] ①37℃ accelerated stability test

[0073] Three batches (LD160613, LD170312, LD170920) of canine distemper detection cards were taken and stored at 37°C. Take three batches of test cards for each measurement. The measurement time interval is the 3rd day, the 7th day, the 10th day, the 14th day, the 17th day and the 20th day. Rate. When the color of the NC film is abnormal in the experimental results and the test results are inaccurate, the detection card is judged to be invalid.

[0074] ②Stability test at room temperature

[0075]Take three batches of test cards (LD160613, LD170312, LD170920) and store them at room temperature. Take a number of three batches of test cards for each measurement. The test time interval is the first month, the third month, and the third month. 6 months, 12 months, 18 months, and 24 months; the content of determination is color development and coincidence rate....

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Abstract

The invention relates to a preparation method of a quantitative detection card for canine distemper virus antigen suitable for the domestic market. The specific steps are: (1) separation, cultivation and purification of canine distemper virus; (2) anti-canine distemper virus pairing Preparation of monoclonal antibody, including immunization procedures, cell fusion, hybridoma screening and cloning, antibody preparation and purification; (3) Canine distemper virus rapid detection colloidal gold detection card and its preparation method, including sample pad treatment, spray film , C, T line determination, the processing of the tested sample, and the performance measurement of the test card. The invention can quickly, sensitively and accurately detect domestic canine distemper virus qualitatively, and overcomes the problem that the paired monoclonal antibody used in the development of domestic canine distemper virus colloidal gold detection card is imported raw material, and the monoclonal antibody of imported raw material is separated from a certain country The current virus preparation overcomes the phenomenon of missed detection due to remote virus mutations in the detection process.

Description

technical field [0001] The invention relates to a detection card suitable for qualitative detection of canine distemper virus antigen in the domestic market and a preparation method thereof, belonging to the technical field of biological detection. Background technique [0002] Canine distemper is an acute, highly contagious viral disease with a higher mortality rate in puppies than any other infectious disease. Canine distemper is also an infectious disease of mink, also known as mink distemper, with high mortality and serious economic losses. For more than 10 years, China's sable market has become larger and larger. In pet stores, we can see a dazzling array of diagnostic products, edible products, and therapeutic products written in English letters. People who love pets trust imports regardless of the price. The product. Even the raw material for the development of antigen diagnostic test strips, monoclonal antibodies, is dependent on imports. For several years, HyTest ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/531G01N33/532
CPCG01N33/56983G01N33/531G01N33/532
Inventor 武玉香于金枝孙珊珊唐世云刘捷
Owner SHANDONG LVDU BIO SICIENCE & TECH
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