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Group A bovine rotavirus detecting kit and preparation method of bovine rotavirus antibody colloidal gold agent

A bovine rotavirus and detection kit technology, applied in the biological field, can solve the problems of high cost of BRoV, affecting the prevention and treatment of BRoV, and achieve the effect of reliable detection results, large market competitiveness, and easy cleaning

Inactive Publication Date: 2018-06-12
SHANDONG VOCATIONAL ANIMAL SCI & VETERINARY COLLEGE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Factors such as high cost, the need for special expensive instruments, the need for special technicians, and the presence of hazardous substances affect the timely diagnosis of BRoV, especially the on-site diagnosis of BRoV, which also affects the timely prevention and treatment of BRoV

Method used

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  • Group A bovine rotavirus detecting kit and preparation method of bovine rotavirus antibody colloidal gold agent

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preparation example Construction

[0032] Wherein, the reagent R2 is a colloidal gold reagent for bovine rotavirus antibody. The invention provides a preparation method of the reagent R2. The method comprises the following steps: step 1) firing colloidal gold; step 2) coupling the antibody with colloidal gold nanoparticles to obtain the gold nanoparticle solution of conjugated antibody; step 3) centrifuge, and resuspend the precipitate with a buffer containing stabilizers and preservatives, and adjust the concentration to is 0.2.

[0033] Among them, step 1) specifically includes: step 1.1) preparing 1% chloroauric acid solution, adding 1 g of chloroauric acid to 100 mL ultrapure water, and storing it in the dark at 4°C for later use; step 1.2) preparing 1% trisodium citrate solution , Dissolve 1g of trisodium citrate in 100mL of ultrapure water and set aside; Step 1.3) Add 100mL of ultrapure water into a cleaned round-bottomed flask, put 1 piece of magnetic stirring bar, add 1mL of 1% chloroauric acid solutio...

specific Embodiment approach

[0039] As a specific embodiment, the preparation method of group A bovine rotavirus detection kit comprises the following steps:

[0040] 1.60nm colloidal gold preparation process

[0041] The required glassware is soaked in potassium dichromate concentrated sulfuric acid lotion for 48 hours, rinsed with tap water, washed with detergent, rinsed with tap water 10 times, rinsed with distilled water 3 times, soaked in distilled water for 24 hours, rinsed with deionized water 3 times, and dried in an oven spare;

[0042] 1% chloroauric acid solution configuration: add 1g chloroauric acid to 100mL ultrapure water, store in the dark at 4°C for later use;

[0043] 1% trisodium citrate solution configuration: 1g trisodium citrate dissolved in 100mL ultrapure water;

[0044] Add 100mL of ultrapure water to a cleaned 1000mL round bottom flask, put a magnetic stirrer, add 1mL of 1% chloroauric acid solution, stir at 600rpm and heat until the solution boils, then quickly add 1% trisodiu...

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Abstract

The invention relates to a group A bovine rotavirus detecting kit. The group A bovine rotavirus detecting kit is composed of an agent R1 and an agent R2. The invention also relates to a preparation method of the group A bovine rotavirus detecting kit. The preparation method of the group A bovine rotavirus detecting kit comprises firing colloidal gold; coupling antibody with colloidal gold nanoparticles to obtain antibody-coupled gold nanoparticle solution; performing centrifugation and settlement, and performing resuspension through buffer solution containing stabilizer and preservatives to regulate the concentration. The group A bovine rotavirus detecting kit is high in detecting sensitivity and specificity, good in stability, simple in operation, short in reaction time, applicable to a fully automatic biochemical analyzer, a special protein analyzer and a spectrophotometer, and capable of avoiding precipitates after reaction and facilitating cleaning of reaction beakers. The group Abovine rotavirus detecting kit is more reliable in detecting results compared with BRoV (bovine rotavirus) detecting data statistical analysis of samples in methods such as IPMA (immunoperoxidase mono-layer assay), ELISA (enzyme-linked immune sorbent assay), immunofluorescence and immunochromatography, and lower in production costs compared with emulsion-process production, thereby being high in marketing competitiveness.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a group A bovine rotavirus detection kit and a preparation method thereof. Background technique [0002] Rotavirus is one of the pathogens of viral diarrhea in various young animals and infants. In 1968, Mebus and others first discovered bovine rotavirus (Bovine rotavirus, BRoV) in the feces of calf diarrhea on a farm in Alaska, USA. Later, bovine rotavirus caused by bovine rotavirus was found in Europe, America, Australia, New Zealand, Japan, etc. calf diarrhea. [0003] BRoV is a double-stranded RNA virus belonging to the genus Rotavirus in the Reoviridae family. BRoV infection causes an acute intestinal infectious disease in calves with intestinal dysfunction, mainly vomiting, diarrhea, dehydration and acid-base balance disorders. characteristics, bring serious harm to the cattle industry. The BRoV virion is slightly round, has a double-layer capsid, and is 65-75nm in diameter....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/531
Inventor 胡士林崔晓辰郑建楠李明群曹雷马爱霞崔尚金
Owner SHANDONG VOCATIONAL ANIMAL SCI & VETERINARY COLLEGE
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