A porcine reproductive and respiratory syndrome virus-swine influenza virus reconstituted virion vaccine and its preparation method and application
A swine flu virus, respiratory syndrome technology, applied in the direction of virus/phage, biochemical equipment and methods, virus, etc., can solve the problem of reduced effect, reduced active ingredients of vaccine, difficult to maintain the immunogenicity and safety of inactivated vaccine, etc. problems, to achieve the effect of enhanced presentation, prolonged vaccine immunization period, and enhanced local and systemic immune responses
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Embodiment 1
[0085] Example 1 Preparation of porcine reproductive and respiratory syndrome virus-swine influenza virus reconstituted virus particles and vaccines
[0086] 1. Virus strain, cell, culture medium
[0087] The vaccine and the PRRSV antibody strain prepared by the invention are the 25th generation virus strain HP-PRRSV-JXM-F25 of the highly pathogenic porcine reproductive and respiratory syndrome virus HP-PRRSV-JXM-F5 strain which is passed on Marc-145 cells. The HP-PRRSV-JXM-F5 strain of the highly pathogenic porcine reproductive and respiratory syndrome virus, which is the adapted strain of HP-PRRSV-JXM uploaded to the 5th generation in MARC-145 cells, is classified and named as the highly pathogenic porcine reproductive and respiratory syndrome virus. Respiratory Syndrome Virus, its microbial deposit number is: CGMCCNO.9453, deposited in the General Microbiology Center of the China Microorganism Culture Collection Administration Committee, the preservation date is July 8, 201...
Embodiment 2
[0151] Example 2 Preparation and quality control method of stable adjuvant complex, porcine reproductive and respiratory syndrome whole virus-swine influenza reconstituted virion vaccine
[0152] 1. Preparation and assay of BCG lysate
[0153] The strain Bacillus Calmette-Guérin (BCG) D2BP302S11 was provided by the China Institute for the Control of Drugs and Biological Products, and the seed batch was established according to the strain management regulations for the production and verification of biological products. It should not exceed 12 generations from the batch of working seeds to the collection of bacterial cells. BCG should float on the surface of Sutong medium, which is a wrinkled and yellow bacterial membrane. The results of virulence test and non-toxic BCG test were negative. Freeze-dried strains are stored at 2-8°C, and liquid strains are stored below -70°C.
[0154] BCG D2BP302S11 was statically cultured at 37°C for 2-3 weeks in modified Sutong comprehensive m...
Embodiment 3
[0184] Example 3 Stability and validity of porcine reproductive and respiratory syndrome intact virus-swine influenza virus reconstituted virion vaccine
[0185] 3.1 Stability of vaccine semi-finished products in liquid frozen state
[0186] The porcine reproductive and respiratory syndrome intact virus-swine influenza virus reconstituted virion vaccine stock solution was prepared according to the method in Example 1, and the stable adjuvant complex and the preparation method of the vaccine antigen were prepared according to the method in Example 2.
[0187] The semi-finished vaccines were stored in polypropylene bottles at -35°C and -70°C, and the inactivated and purified porcine reproductive and respiratory syndrome intact virus-swine influenza virus reconstituted virion particles were detected every 3 months. The results are shown in Table 1 below.
[0188] Table 1 Stability of semi-finished vaccines stored in frozen state
[0189]
[0190]
[0191] The results in Ta...
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