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Detection card for rapidly detecting canine distemper virus antigens and preparation method of detection card

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 being suitable for popularization, efficient and sensitive detection, and high sensitivity

Active Publication Date: 2019-07-05
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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  • Detection card for rapidly detecting canine distemper virus antigens and preparation method of detection card
  • Detection card for rapidly detecting canine distemper virus antigens and preparation method of detection card
  • Detection card for rapidly detecting canine distemper virus antigens and preparation method of detection card

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: a kind of preparation method of rapid detection canine distemper virus pathogen detection card, concrete steps are:

[0038] (1) Isolation, cultivation and purification of canine distemper virus.

[0039] Select Shandong, Guangdong, Sichuan and other 3 regions, and the ocular secretions diagnosed as strong positive by PCR were inoculated into F81 cells for 3 generations, and the cells with significant lesions were collected, frozen, expanded and cultured, collected for poisoning, and concentrated by ultrafiltration 10 times, then repeated freeze-thaw 3 times, centrifuged at 10000rpm for 30 minutes, applied sucrose density gradient, 30%, 35%, 40%, 45% 4 gradients, centrifuged at 26000rpm for 30 minutes, took out each layer, and determined the location and location of the virus. content.

[0040] (2) Preparation of paired monoclonal antibodies against canine distemper virus

[0041] ①Immunization program

[0042] Mix the purified virus from three regions ...

Embodiment 2

[0062] Embodiment 2: Pretreatment method and detection method of tested sample

[0063] The using method of canine distemper virus pathogen rapid detection card of the present invention is as follows:

[0064] a. Use a cotton swab to collect dog, mink, fox eye secretions or nasal fluid, saliva.

[0065] b. Open the sample collection tube and insert the swab.

[0066] c. Agitate the swab well until the sample on the swab dissolves into the diluent, then discard the swab.

[0067] d. After oscillating and mixing, vertical the sample tube, absorb the supernatant and drop 3-5 drops into the sample hole in the test card.

[0068] e. The result is judged at 5 minutes, and the judgment at other times is invalid.

[0069] (1) Positive ( figure 2 ): If the C line shows color and the T line shows color, it is judged as positive.

[0070] (2) Negative ( image 3 ): If the C line shows color and the T line does not show color, it is judged as negative.

[0071] (3) invalid ( Fi...

Embodiment 3

[0072] Embodiment 3: Test card performance measurement

[0073] (1) Stability

[0074] ① Accelerated stability test at 37°C

[0075] Several batches of canine distemper test cards (LD160613, LD170312, LD170920) were taken and stored at 37°C. Three batches of test cards were taken for each measurement, and the measurement time intervals were 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 and the test result is inaccurate in the experimental results, it is determined that the test card is invalid.

[0076] ② Stability test at room temperature

[0077] Take three batches of test cards (LD160613, LD170312, LD170920) and store them at room temperature. Take three batches of test cards for each measurement. The test time interval is the first month, the third month, and the third month. 6th month, 12th month, 18th month, and 24th month; the measurement content is color development and coinciden...

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Abstract

The invention relates to a detection card for rapidly detecting canine distemper virus antigens and a preparation method of the detection card. The method includes the following specific steps of: (1)the isolation, culture and purification of canine distemper viruses; (2) the preparation of paired monoclonal antibodies against the canine distemper viruses, wherein the preparation involves animmunization procedures, cell fusion, hybridoma screening and cloning, antibody preparation and purification; and (3) the preparation method of the colloidal gold detection card for rapidly detecting the canine distemper viruses, wherein the preparation method involves sample pad treatment, film spraying, and C, T line determination, to-be-tested sample processing and test card performance measurement.With the preparation method of the detection card of the invention adopted, canine distemper viruses in China can be quickly, sensitively and accurately detected. According to the prior art, paired monoclonal antibodies for developing colloidal gold detection cards for detecting canine distemper viruses in China are made of imported raw materials; the imported raw materials of the monoclonal antibodies are prepared from current viruses isolated by a certain country. With the preparation method of the invention adopted, the conditions in the prior art can be avoided, and a phenomenon that missed detection caused by virus variation due to long distances between areas during a detection process can be avoided.

Description

technical field [0001] The invention relates to a detection card for qualitative detection of canine distemper virus antigen suitable for 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, and the mortality rate of this disease in young dogs is higher than any other infectious disease. Canine distemper is also an infectious disease of mink, also known as mink distemper, with a high mortality rate and serious economic losses. For more than 10 years, China's dog and mink market has been growing. In pet stores, we can see a wide variety 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 monoclonal antibody, the raw material for the development of antigen diagnostic test strips, is still dependent on im...

Claims

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Application Information

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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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