Colloidal gold detection card for quickly detecting rabbit hemorrhagic disease virus (RHDV) and preparation method thereof

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

Active Publication Date: 2019-06-07
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 dete

Method used

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  • Colloidal gold detection card for quickly detecting rabbit hemorrhagic disease virus (RHDV) and preparation method thereof
  • Colloidal gold detection card for quickly detecting rabbit hemorrhagic disease virus (RHDV) and preparation method thereof
  • Colloidal gold detection card for quickly detecting rabbit hemorrhagic disease virus (RHDV) and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: A preparation method of colloidal gold test 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 one of the anti-rabbit plague virus VP60 hybridoma cell lines, 2F4, which was deposited in the China 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 results, the company continues to prepare hybridoma cell lines paired with it, using the following methods to prepare:

[0035] ①Immunization program:

[0036] Insect cell expression protein VP60 immunogen (Shandong Binzhou Animal Husbandry and Veterinary Research Institute) and 501 adjuvant (Shandong Lvdu Biotechnology Co., Ltd., article number LD003) were mixed at a volume ratio of 1:0.2-0.3, for 8-10 weeks Immunization of Balb / c mice of the age of 50μg / mouse into the leg muscles, and then...

Embodiment 2

[0056] Embodiment 2: Pre-processing method and detection method of tested sample

[0057] Take the liver and kidney of the newly deceased rabbit, weigh 5g, add 5ml sample diluent, grind to paste, freeze-thaw repeatedly 3 times, then centrifuge at 12000rpm for 5 minutes, take the supernatant for testing.

[0058] The method of using the rapid detection card for the rabbit plague virus pathogen of the present invention is to first process the sample according to the sample pre-processing method, then sample 200 microliters of the extract and add it to the sample hole of the detection card, and determine the result within 5-10 minutes . The result is judged 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 as negative.

[0061] (3) invalid ( Figure 4 ): In the observation hol...

Embodiment 3

[0062] Example 3: Test card performance measurement

[0063] (1) Stability test

[0064] ①37℃ accelerated stability test:

[0065] Take three batches (LD160217, LD160312, LD160525) of rabbit plague detection cards and store them at 37°C. Take three batches of test cards for each measurement. The measurement interval is the 3rd day, the 7th day, the 10th day, the 14th day, the 17th day, and the 20th day. The measurement content is the color development and compliance rate. When the color of the NC film is abnormal in the experiment result and the test result is inaccurate, it is determined that the test card is invalid.

[0066] ②Stability test at room temperature

[0067] Take three batches of test cards (LD160217, LD160312, LD160525) 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 first month. 6 months, 12th month, 18th month, 24th month; the content of the measurement is ...

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Abstract

The invention relates to a colloidal gold detection card for quickly detecting rabbit hemorrhagic disease virus (RHDV) and a preparation method thereof. The method comprises the following steps: 1) preparation of paired monoclonal antibodies against RHDV, wherein the preparation processes comprise immune procedure, cell fusion, hybridoma screening and cloning and antibody preparation and purification; and 2) preparation of the colloidal gold detection card for quickly detecting rabbit hemorrhagic disease virus (RHDV), wherein the preparation processes comprise preparation of a colloidal gold monoclonal antibody, sample pad treatment, membrane spraying, determination of C and T lines, treatment of tested samples and performance determination. The method can quickly, sensitively and accurately detect RHDV infection in the liver and kidney of a rabbit, and overcomes the problem that erythrocyte agglutination test is the main method to detect the RHDV in China, thereby greatly reducing detection time and providing convenience for on-site detection.

Description

Technical field [0001] The invention relates to a colloidal gold detection card for rapidly detecting rabbit plague virus and a preparation method thereof, and belongs 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 is the most serious threat to rabbits. Its outstanding characteristics are rapid onset, rapid transmission, short time from onset to death, and high mortality. high. Once a herd of rabbits is infected, it is often annihilated. This disease has the highest incidence in young rabbits and adult rabbits, and is not affected by the season. Generally, it is most likely to break out in spring and autumn. Rabbit farmers must strictly prevent and control rabbit plague, otherwise, once contracted with this disease, it will inevitably cause very significant economic losses. Rabbit plague virus (RHDV) infection will deal a devastating blow to the rab...

Claims

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Application Information

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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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