A porcine circovirus type 3 virus strain and its application
A technology of porcine circovirus and type 3 virus, applied in antiviral agents, virus/bacteriophage, virus antigen components, etc., can solve problems such as vaccine immunization failure
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0030] Example 1: Isolation and identification of porcine circovirus type 3 ZM-PCV3-14CGMCC No.17290.
[0031] 1.1 Experimental materials
[0032] The disease samples used in the experiment came from pig farms with porcine circus disease in Guangdong Province, and the collection time was January 2018; the porcine kidney cell line (Porcine Kidney, PK-15) was purchased from the American Culture Center ATCC.
[0033] 1.2 Primer design
[0034]Referring to the PCV3KY965242.1 gene series published in GenBank, two pairs of specific primers were designed using Primer 5, and the primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. The nucleotide sequence of the specific primer is as follows:
[0035] PCV3-F-1: 5'-TAGTATTACCCGGCACCTC-3';
[0036] PCV3-R-1: 5'-GCCCGGCACCAAAATGAGAC-3';
[0037] PCV3-F-2: 5'-TTAGAGAACGGACTTGTAAC-3';
[0038] PCV3-R-2: 5'-ACAGCTGGCACATACTACAC-3';
[0039] 1.3 Collection and processing of disease materials
[0040] Collect samples o...
Embodiment 2
[0057] Embodiment 2: the mensuration of porcine circovirus type 3 virus strain ZM-PCV3-14 virus titer
[0058] The isolated virus liquid was inoculated on the PK15-B1 cell monolayer, adsorbed at 37°C for 30 minutes, added DMEM cell maintenance solution containing 2% calf serum, cultured at 37°C for 60 hours, continued to pass to the 20th passage, and stored at -20°C. Use DMEM cell maintenance solution to make 10 for PCV3 -1 ~10 -8 Serially dilute and inoculate a monolayer of PK15-B1 cells in a 96-well plate, inoculate 4 wells for each dilution, and inoculate 100 μl in each well. At the same time, a negative control was set up, placed in a CO2 incubator for 12 hours at 37° C., treated with 300 mM D-glucosamine hydrochloride, and incubated at 37° C. for 48 hours. Fix the cells with absolute ethanol, measure the number of wells containing green fluorescent substance cells in each dilution by the IFA method, and finally calculate the TCID of the virus by the Reed-Muench method ...
Embodiment 3
[0059] Embodiment 3: the preparation of porcine circovirus type 3 ZM-PCV3-14 strain inactivated vaccine
[0060] 3.1 Virus culture
[0061] Use low-sugar DMEM medium containing 10% fetal bovine serum, 100U / ml penicillin, 100g / ml streptomycin, T225cm 2 Cell culture flask, 37°C, 5% CO 2 Cultivate PK-15 cells adherently in an incubator; when the fusion degree reaches about 80%, discard the culture medium, inoculate porcine circovirus type 3 ZM-PCV3-14 strain, and then use 2% fetal bovine serum, 100U / ml penicillin, 100g / ml streptomycin low-sugar DMEM medium, cultured at 37°C, 5% CO2 for 12h, treated with 300mM D-glucosamine hydrochloride, continued to culture at 37°C for 48h, and harvested the virus culture; freeze-thawed repeatedly Centrifuge at 10,000×g for 10 minutes three times to remove cell debris and collect the supernatant to obtain porcine circovirus type 3 virus liquid, which is stored at -80°C for later use.
[0062] 3.2 Inactivation of virus liquid and inactivation...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com