Latex agglutination detection method for bovine viral diarrhea viruses and application

A bovine viral diarrhea and detection method technology, applied in the fields of immunology and chemistry, to achieve high sensitivity, good specificity and reactogenicity, and strong specificity

CN112034165AInactive Publication Date: 2020-12-04NINGXIA UNIVERSITY
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2020-12-04
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention provides a bovine viral diarrhea virus antibody detection method which uses a latex agglutination test method and application. Prokaryotic expression purified product E0 protein is coupled to carboxylated latex microspheres by using carbodiimide, so that a rapid detection method for bovine viral diarrhea, namely a latex agglutination detection method is established. The method has the advantages of simplicity and convenience in operation, low requirements on equipment, strong specificity, high sensitivity, rapidness in detection and the like.
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Description

technical field

[0001] The invention belongs to the technical field of immunology, and relates to the related fields of immunology and chemistry. Specifically, it relates to a latex agglutination detection method and application of bovine viral diarrhea virus antibody. Background technique

[0002] Bovine viral diarrhea virus (BVDV) is a member of the genus Pestivirus of the family Flaviviridate and can cause bovine viral diarrhea, also known as bovine viral diarrhea / mucosal disease (Bovine viral diarrhea-mucosal disease, BVD-MD). The virus can cause viral diarrhea in cattle and other ruminants, seriously affect the reproductive system, digestive system, immune system and milk production, cause huge economic losses to the breeding industry, and affect the development of the animal husbandry industry. Research surveys in my country have shown that more than 46.7% of cattle farms are positive for bovine viral diarrhea virus antigens, and 2.2% of the cattle in the herd are pe...

Examples

Embodiment Construction

[0012] If the specific experimental conditions and methods are not indicated in the examples of the present invention, conventional conditions are generally followed.

[0013] 1. Induced expression of recombinant plasmid pET-32a-E0: Take a small amount of BL21 bacterial liquid containing pET-32a-E0 with a disposable sterile inoculation stick, and inoculate it on LB solid medium (containing 100 μg / mL Amp) Separation culture by streaking, placed in a constant temperature incubator at 37°C, 12~15h. Dip a sterile inoculation stick to pick up a single colony into 5mL LB liquid medium (containing 100μg / mL Amp), place it in a horizontal water bath shaker at 37°C, and culture overnight for 12-15h. Absorb the overnight cultured bacterial liquid into the freshly prepared LB liquid medium (containing 100 μg / mL Amp) at a ratio of 1:50, incubate on a shaker (220 r / min) at 37°C for 2 to 3 hours, then take an appropriate amount of bacterial liquid Measure OD with UV spectrophotometer 600 T...