Canine distemper virus colloidal gold immunochromatographic test strip and preparation method thereof

A technology for immunochromatographic detection and canine distemper virus, which is applied to measuring devices, analytical materials, instruments, etc., can solve the problems of cumbersome and time-consuming detection steps, and achieve the effects of easy promotion, simple and fast operation, and high sensitivity

Inactive Publication Date: 2016-12-14
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problems that existing CDV detection technology has cumbersome and time-consuming detection steps, requires professionals to operate and mainly relies on laboratories, the present invention provides a colloidal gold immunochromatographic detection test strip for canine distemper virus and its Preparation

Method used

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  • Canine distemper virus colloidal gold immunochromatographic test strip and preparation method thereof
  • Canine distemper virus colloidal gold immunochromatographic test strip and preparation method thereof

Examples

Experimental program
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Embodiment 1

[0033] The preparation of embodiment 1 canine distemper virus monoclonal antibody N18, C42

[0034] (1) The purified canine distemper virus was inoculated into Balb / c mice, and after booster immunization, the splenocytes were collected, fused with myeloma cells prepared in advance, and cultured with 1% HAT medium; the fusion was about 2 weeks, Two positive hybridoma cell lines were screened out by indirect ELISA method, and the screened positive hybridoma cell lines were cloned to obtain two canine distemper virus monoclonal antibody hybridoma cell lines;

[0035](2) Choose female Balb / c mice aged 6-8 weeks, intraperitoneally inject pristane 0.5mL / only, and intraperitoneally inject two strains of canine distemper virus monoclonal antibody hybridoma cells respectively after 7 days, and the inoculation amount is 4 ×10 6 ~6×10 6 Cells / mL / mouse; After the mouse abdomen swells (about one week), extract the ascites, centrifuge at 5000rpm for 15 minutes, and take the supernatant; s...

Embodiment 2

[0036] The preparation of embodiment 2 colloidal gold solution

[0037] Colloidal gold solution was prepared by trisodium citrate reduction method: take 1% chloroauric acid solution and add 99ml of ultrapure water to configure a final concentration of 0.01% chloroauric acid solution. After heating to boiling, add 1% citric acid trisodium Add 1ml of sodium and continue heating, the solution turns from light yellow to blue-black and finally wine red. After the color is stable, continue heating for 5 minutes, cool at room temperature and store at 4°C for later use to obtain a colloidal gold solution. Take a small amount of colloidal gold particles and observe them with a transmission microscope. The size of the colloidal gold particles is basically the same and the distribution is uniform. The diameter of the colloidal gold particles is about 40nm.

Embodiment 3

[0038] Example 3 Preparation of canine distemper virus monoclonal antibody N18 and colloidal gold coupling label (gold-labeled canine distemper virus monoclonal antibody N18)

[0039] (1) Stepwise dilution method determines the consumption ratio of canine distemper virus monoclonal antibody N18 and colloidal gold solution: adjust the pH value of colloidal gold solution with 0.1mol / L potassium carbonate solution to be 8.4, get 11 clean centrifuge tubes , numbered from tube No. 1 to tube No. 11, add 1ml of colloidal gold solution to each tube; the contents of purified canine distemper virus monoclonal antibody N18 are 2 μg, 4 μg, 6 μg, 8 μg, 10 μg, 12 μg, Add 14μg, 16μg, 18μg, 20μg to tube No. 2 to tube No. 11 according to the above order, and mix with the colloidal gold solution in the centrifuge tube. After 5 minutes, add 10% to tube No. 2 to tube No. 11 Mix 1ml of the sodium chloride solution, and let it stand at room temperature for more than 2 hours to observe the results; ...

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Abstract

The invention provides a canine distemper virus colloidal gold immunochromatographic test strip and a preparation method thereof, relates to the technical field of colloidal gold test strip, and solves the problems of complex test procedures, time consumption, professional operation requirement and dependence on the laboratory in the existing CDV detection techniques. The test strip includes a PVC base plate, and a sample pad, a gold-labeled combined pad, a nitrocellulose membrane with a detection line and control line, and an absorption pad, which are pasted on the base plate. The gold combined pad is wrapped with a coupling marker of purified canine distemper virus monoclonal antibody N18 and colloidal gold; the detection line on the nitrate cellulose membrane is wrapped with a purified canine distemper virus purified monoclonal antibody C42; and the control line on the nitrocellulose membrane is coated with a goat-anti-mouse IgG. The test strip of the invention has the advantages of simple and rapid operation, clear and easy detection, strong specificity, high sensitivity, and no need of instruments and equipment.

Description

technical field [0001] The invention relates to the technical field of colloidal gold detection test strips, in particular to a colloidal gold immunochromatographic detection test strip for canine distemper virus and a preparation method thereof. Background technique [0002] Canine distemper (Canine Distemper, CD) is a highly contagious, fatal infectious disease caused by Canine Distemper Virus (CDV). The clinical symptoms of fur animals infected with canine distemper virus are mental depression, elevated body temperature, anorexia, and diarrhea. In the later stage, neurological symptoms appear, manifested as ataxia and quadriplegia. In the end, he foamed at the mouth, screamed and died. Known as one of the three major diseases of fur animals. [0003] Existing canine distemper virus detection methods mainly include serum neutralization test, immunofluorescence test, ELISA, RT-PCR, virus isolation and inclusion body inspection. However, these methods require professional...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/531G01N33/532G01N33/543
CPCG01N33/56983G01N33/531G01N33/532G01N33/54306
Inventor 易立曹智刚程悦宁仝明薇王建科林鹏程世鹏闫喜军
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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