Vaccine composition containing porcine circovirus type 2 antigen and swine influenza antigen
A technology of porcine circovirus and vaccine composition, which is applied in the field of pig vaccines, and can solve problems such as the impact of pig immune function, no prevention and treatment of swine influenza virus disease, and reduced growth performance of pigs
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Embodiment 1
[0103] Example 1. Isolation and Identification of Swine Influenza Strains
[0104] The isolation and identification of SIVH1N1 strain of the present invention and SIVH3N2 strain:
[0105] SIVH1N1 subtype virus (named ZJS strain) and SIVH3N2 subtype virus (named WX strain) were identified by RT-PCR identification, hemagglutination test and animal inoculation test.
[0106] SIVRT-PCR identification primers
[0107]
[0108] Such as figure 1 As shown, use the above SIVRT-PCR identification primers to identify SIVH1N1 and SIVH3N2: Lane M is Marker, A in lane 1 is the amplified band of HA gene of SIVH1N1 subtype ZJS strain, and B in lane 2 is SIVH1N1 subtype ZJS The amplified band of the NA gene of the strain, A in lane 3 is the amplified band of the HA gene of the SIVH3N2 subtype WX strain, and B in lane 4 is the amplified band of the NA gene of the SIVH3N2 subtype WX strain. The results show Both the HA gene and NA gene of the two viruses could amplify specific bands.
[...
Embodiment 2
[0116] Example 2: Preparation of porcine circovirus type 2 antigen and swine influenza virus antigen
[0117] Preparation of porcine circovirus type 2 (SH strain) virus liquid: the PCV2SH strain seed was mixed with 0.1TCID 50 The inoculum amount per cell was inoculated on a monolayer of PK15 (purchased from ATCC, model ATCCCCL-33 TM ) cells, adsorb at 37°C for 30 minutes, add MEM liquid medium containing 4% (volume ratio) calf serum and 2mMD-glucosamine hydrochloride (prepared with MEM dry powder medium purchased from Life Technologies, USA, according to its instructions), Place at 37°C to continue culturing. Observe twice a day, the cells grow well, harvest the cell culture after 4 days of culture at 37°C, freeze and thaw 3 times, harvest the virus, and use PBS to dilute the harvested virus liquid to a virus titer of 10 6.0 TCID 50 / mL, the batch number is 20110428.
[0118] Preparation of porcine circovirus type 2 (SH strain) subunit antigen: clone porcine circovirus typ...
Embodiment 3
[0121] Embodiment 3: treatment and inactivation of virus fluid or bacteria fluid.
[0122] Treatment of porcine circovirus type 2 (SH strain) virus liquid: filter the virus liquid prepared in Example 2 with batch number 20110428 through a hollow fiber (0.5 μm ~ 2 μm) filter column (purchased from GE Healthcare Life Sciences) to remove cell debris, and then add Formaldehyde solution with 0.2% to 0.3% virus volume was inactivated at 37°C for 18 hours. Porcine circovirus type 2 whole virus antigen was obtained, and the antigen content was that the virus titer before inactivation was 10 6.0 TCID 50 / ml, the batch number is 20110506.
[0123] Treatment of swine influenza virus liquid: filter the H1N1 subtype SIV antigen liquid with batch number 20110502 in Example 2 with a hollow fiber (0.5 μm ~ 2 μm) filter column (purchased from GE Healthcare Life Sciences) to remove cell debris, and then add 0.2% ~ 0.3% formaldehyde solution was inactivated at 37°C for 18 hours, and the batch...
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