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Rabies virus IgG antibody immune gold-labeled test paper and preparation method thereof

A rabies virus and detection test paper technology, which is applied in the field of rabies virus IgG antibody immunogold standard detection test paper and preparation, can solve the problem of no rabies virus IgG antibody, etc., and achieves the effects of high sensitivity, simple operation and strong specificity

Inactive Publication Date: 2015-02-18
CHANGCHUN SR BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, immune colloidal gold test strips have been widely used in medical and food testing and other fields, such as: AIDS, hepatitis, etc., but the application of this technology in all aspects of animal medicine is still in its infancy, and it has not been used to detect immune animals. Rabies virus IgG antibody level immune gold standard detection test paper product

Method used

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  • Rabies virus IgG antibody immune gold-labeled test paper and preparation method thereof
  • Rabies virus IgG antibody immune gold-labeled test paper and preparation method thereof
  • Rabies virus IgG antibody immune gold-labeled test paper and preparation method thereof

Examples

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Embodiment 1

[0030] The purification of embodiment 1 rabies virus antigen

[0031] Rabies virus was inoculated into monolayer BSR cells at MOI=0.01, 5%CO 2 After culturing at 33°C for 4-5 days, the cultured cell supernatant was harvested twice every 2 days. Add β-propiolactone at a ratio of 1:4000 to the virus volume, shake well, and keep at 4°C for 24 hours to inactivate rabies virus. Place it at 37°C for 2h to hydrolyze β-propiolactone. The inactivated rabies virus was centrifuged at 3,000 rpm for 30 min at 4°C to remove cell debris, and the supernatant was collected for zinc acetate precipitation. Add 1M zinc acetate to the supernatant according to 1 / 50 volume, shake well while adding, let stand at 4°C for 30-60min, centrifuge at 3000rpm at 4°C for 30min, discard the supernatant, and use 1 / 100 of the original supernatant volume for the precipitation to saturate Suspend in EDTA and dissolve overnight at 4°C. Centrifuge at 3000 rpm at 4°C for 30 minutes to remove undissolved substance...

Embodiment 2

[0033] Example 2 Preparation of immunized animal rabies virus IgG antibody immune gold standard detection test paper

[0034] Preparation of serum-treated pad: fully soak the glass cellulose membrane in 20 mM phosphate buffer containing 0.05M tetraborate, 1% bovine serum albumin, 0.04% sodium azide, 0.2% Tween-20 solution, take it out and dry it at 37°C.

[0035] Firing of 20-30nm colloidal gold: In a fume hood, immerse a 250ml Erlenmeyer flask in chloroform siliconizing solution of 5% dichlorodimethylsilane, and then slowly rotate the vessel so that the inner wall of the glassware can be soaked in the siliconizing solution. Let it stand for 10 minutes, take out the glassware, rinse it with a large amount of double distilled water, cover the mouth of the bottle with tinfoil and set aside. Take the siliconized Erlenmeyer flask, add 99mL of double distilled water and 1mL of 1% chloroauric acid, put it on a magnetic heating stirrer, heat to boiling, quickly add 1.6mL of 1% triso...

Embodiment 3

[0040] Embodiment 3 Evaluation of immune gold standard detection test paper for immune animal rabies virus IgG antibody

[0041] Use of immunocolloidal gold detection test paper and result judgment: use sample diluent (phosphate buffer saline solution of 20mM pH 8.0 containing 0.2% Tween-20, 0.04% sodium azide) to carry out 100-fold dilution of the serum sample to be tested (wherein, serum Sample 1 is healthy dog ​​serum with high rabies IgG antibody level confirmed by fluorescent antibody virus neutralization test (FAVNT), serum sample 2 is healthy dog ​​serum with low rabies IgG antibody level confirmed by FAVNT test, serum sample 3 is Serum from healthy dogs without rabies IgG antibodies confirmed by FAVNT test). Put the test paper flat on the table, take 50 μL of the diluted serum sample and add it to the serum processing pad. After standing at 25°C for 15 minutes, judge the result according to the following conditions: the test paper is valid: a purple-red line appears o...

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Abstract

The invention relates to rabies virus IgG antibody immune gold-labeled test paper. The test paper consists of a serum treatment pad, a gold-labeled release pad, a nitrocellulose membrane, an absorbent pad and a back plate, wherein the serum treatment pad, the gold-labeled release pad, the nitrocellulose membrane and the absorbent pad are sequentially superposed and adhered to the back plate, and the superposition distance between every two parts is 2-3mm; a purified rabies virus antigen-colloidal gold compound is coated on the gold-labeled release pad; a detection line and a quality control line are coated on the nitrocellulose membrane; the detection line is fixedly provided with SPA; and the quality control line is fixedly provided with anti-dog rabies virus positive IgG. The test paper disclosed by the invention is easy to operate, high in specificity and high in sensitivity, special instruments and equipment and professionals are not needed, whether vaccine immunity animals generate antibodies on a protection level can be rapidly screened, and a reference basis is provided for vaccine immunity and immune procedure formulation of rabies.

Description

technical field [0001] The invention relates to the technical field of rapid detection, in particular to a rabies virus IgG antibody immunogold standard detection test paper and a preparation method thereof. Background technique [0002] Rabies is an important zoonotic infectious disease caused by Rabies virus (RABV), with a fatality rate of almost 100%. It is one of the major infectious diseases that threaten the health of the population and public health in my country. More than 90% of human rabies is transmitted by licking or biting infected or infected animals. According to incomplete statistics, the proportions of human rabies caused by different kinds of animals in my country are: 85%-95% for dogs, 4% for cats, and 3% for wild animals. At present, there is no effective drug for the treatment of rabies, and once the onset occurs, almost 100% of them die. The key measures to prevent and control rabies are still to strengthen the immunity of dogs, cats and other transmi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/56983G01N2333/145
Inventor 金宏丽夏晓红夏振强石晶王慧慧陈宪平刘冰付玉杨佳
Owner CHANGCHUN SR BIOLOGICAL TECH
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