A colloidal gold detection card for rapid detection of rabbit plague virus and its preparation method

A technology of rabbit distemper virus and colloidal gold, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of ineffective detection, difficulty in RHDV detection, and insufficient detection methods of rabbit distemper virus, and achieves convenient on-site detection and is suitable for promotion The effect of using and shortening the detection time

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

However, this method cannot effectively detect whether the seropositivity is caused by viral infection or seropositivity after vaccination
[0005] Based on the problem that the existing detection methods of rabbit distemper virus are not abundant, the market needs effective detection methods of rabbit distemper virus to solve the technical problem of RHDV detection difficulty

Method used

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  • A colloidal gold detection card for rapid detection of rabbit plague virus and its preparation method
  • A colloidal gold detection card for rapid detection of rabbit plague virus and its preparation method
  • A colloidal gold detection card for rapid detection of rabbit plague virus and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1: A preparation method of a colloidal gold detection card for rapid detection of rabbit plague virus, the specific steps are: (1) Preparation of anti-rabbit plague virus VP60 monoclonal antibody

[0034] The company has obtained 2F4, one of the anti-rabbit plague virus VP60 hybridoma cell lines, which was deposited in the China Center for Type Culture Collection on June 11, 2015, with the deposit number CCTCC NO: C201582, and the deposit address is Wuhan-Wuhan University, China. Based on the above achievements, the company continues to prepare the hybridoma cell lines that are paired with it, using the following methods to prepare:

[0035] ①Immunization program:

[0036] The insect cell-expressed protein VP60 immunogen (Binzhou Animal Husbandry and Veterinary Research Institute, Shandong Province) and 501 adjuvant (Shandong Lvdu Biotechnology Co., Ltd., product number LD003) were mixed in a volume ratio of 1:0.2-0.3 for 8-10 weeks. Balb / c mice were immuniz...

Embodiment 2

[0056] Embodiment 2: Pretreatment method and detection method of tested sample

[0057] The liver and kidney of the newly deceased rabbit were taken, weighed 5 g, added 5 ml of sample diluent, ground to a paste, repeated freezing and thawing 3 times, and then centrifuged at 12,000 rpm for 5 minutes, and the supernatant was collected for detection.

[0058] The method of using the rapid detection card for the pathogen of rabbit distemper virus of the present invention is to first process the sample according to the sample pretreatment method, then take 200 microliters of the sample extract and add it to the sample hole of the detection card, and determine the result within 5-10 minutes. . The result is determined as figure 2 , 3 and 4.

[0059] (1) Positive ( figure 2 ): If the C line develops color and the T line develops color, it is judged as positive.

[0060] (2) negative ( image 3 ): If the C line develops color and the T line does not develop color, it is judged...

Embodiment 3

[0062] Example 3: Test Card Performance Measurement

[0063] (1) Stability test

[0064] ①37℃ accelerated stability test:

[0065] Take a number of rabbit plague detection cards from three batches (LD160217, LD160312, LD160525) and store them 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.

[0066] ②Stability test at room temperature

[0067] Take three batches of test cards (LD160217, LD160312, LD160525) 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 measurement content is color development and coinci...

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Abstract

The invention relates to a colloidal gold detection card for rapidly detecting rabbit plague virus and a preparation method thereof. The specific steps are: (1) preparation of paired anti-rabbit plague virus monoclonal antibodies, including immune procedures, cell fusion, hybridoma screening and Cloning, antibody preparation and purification; (2) colloidal gold detection card for rapid detection of rabbit plague virus and its preparation method, including preparation of colloidal gold monoclonal antibody, sample pad treatment, spray film, C and T line determination, tested Sample processing, performance measurement. The invention can quickly, sensitively and accurately detect rabbit plague virus infection in the liver and kidney of rabbits, overcomes the domestic method of detecting rabbit plague virus mainly by red blood cell agglutination test, greatly shortens the detection time, and provides convenience for on-site detection.

Description

technical field [0001] The invention relates to a colloidal gold detection card for rapid detection of rabbit plague virus and a preparation method thereof, belonging to the technical field of biological detection. Background technique [0002] Rabbit plague, also known as rabbit viral hemorrhagic syndrome, is a viral infectious disease that poses the most serious threat to rabbits. high. Once a rabbit herd is infected, the herd is often wiped out. The disease has the highest incidence in young rabbits and adult rabbits, and is less affected by seasons, generally in spring and autumn. Rabbit farmers must strictly prevent and control rabbit plague, otherwise, once infected with this disease, it will inevitably bring very significant economic losses. Rabbit Distemper Virus (RHDV) infection will give a devastating blow to the rabbit population, which is extremely harmful. RHDV is one of the viruses that must be checked for rabbits in the rabbit industry and laboratory use. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/531G01N33/532
Inventor 武玉香于金枝曲光刚张莎莎
Owner SHANDONG LVDU BIO SICIENCE & TECH
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