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Porcine circovirus II type-porcine pseudorabies double-combination vaccine, and preparation methods and application thereof

A technology of porcine circovirus and porcine pseudorabies, which is applied in the field of vaccines, can solve the problems of serious losses and aggravation of piglets, and achieve the effects of reducing the number of vaccinations, enhancing the immune effect, and reducing costs

Inactive Publication Date: 2014-07-16
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, studies have found that PCV2 and PRV have a synergistic effect when co-infected, leading to aggravated disease and more serious losses in piglets after weaning

Method used

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  • Porcine circovirus II type-porcine pseudorabies double-combination vaccine, and preparation methods and application thereof
  • Porcine circovirus II type-porcine pseudorabies double-combination vaccine, and preparation methods and application thereof
  • Porcine circovirus II type-porcine pseudorabies double-combination vaccine, and preparation methods and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The preparation of embodiment 1 porcine circovirus type Ⅱ antigen

[0030] 1 Preparation of porcine circovirus type Ⅱ subunit antigen

[0031] (1) Introduce the porcine circovirus type 2 ORF2 gene into the melittin signal peptide gene sequence, transfer it into the pMD-18T vector, and obtain a recombinant vector containing the porcine circovirus type 2 ORF2 gene: first design the primer Spe-ATG(5' -cgactagtatgacgtatccaaggaggcgt-3') and primer Hind-TAA (5'-gcaagctttatcattaagggttaagttggg-3'), using the genomic DNA of the PCV2-SH strain with the preservation number CGMCC No.2389 as a template to amplify the PCV2-ORF2 gene, and amplify the The increased ORF2 gene was cloned into the pMD-18T vector to obtain the recombinant vector pT-ORF2. The size of the PCR product is about 730bp. The PCR product was sequenced. According to the results of electrophoresis and sequencing, the recombinant vector pT-ORF2 contained the sequence of SEQ ID NO1 in the sequence list. For the sp...

Embodiment 2

[0045] The preparation of embodiment 2 porcine pseudorabies virus antigen

[0046] The porcine pseudorabies virus gene deletion strain (SA215 strain, TK ˉ / gE ˉ / gI ˉ The deletion of three genes is disclosed in Chinese patent CN101186902, the strain is deposited in CCTCC with the preservation number V200002) is properly diluted with virus diluent (serum-free DMEM medium), and inoculated in ST cells ( (purchased from CCTCC, No. GDC0060), cultivated at 37°C for 30 min, added DMEM cell maintenance solution containing 3% (v / v) calf serum, cultured at 37°C for 2-4 days, freeze-thawed 2-3 times, and harvested the virus. Virus titer is 10 8.7 TCID 50 / ml.

Embodiment 3

[0047] Example 3 Preparation of porcine circovirus type Ⅱ antigen and porcine pseudorabies virus antigen dual vaccine

[0048] 1. Porcine circovirus type Ⅱ inactivated vaccine diluted porcine pseudorabies freeze-dried live vaccine

[0049] Mix the inactivated antigen of porcine circovirus type Ⅱ whole virus with the water quality adjuvant IMS1313N VG at a ratio of 1:1, stir and mix with an IKA mixer (350rpm) for 5 minutes to make an emulsion. It is tested according to the national standard and all meet the regulations. The specific vaccine ratio is carried out with reference to Table 1. There are 7 kinds of A to G in total. Among them, the porcine circovirus type Ⅱ in groups A, B, C, D, E, and F uses PCV2 whole virus inactivated antigen, and each head (2ml ) in the content of 10 3.0 TCID 50 、10 4.0 TCID 50 、10 6.0 TCID 50 、10 6.0 TCID 50 、10 7.0 TCID 50 and 10 6.0 TCID 50 .

[0050] A, B, C, D, E, G groups of porcine pseudorabies virus antigen and lyoprotectant...

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Abstract

The invention relates to a porcine circovirus II type-porcine pseudorabies double-combination vaccine which is characterized by including at least one porcine circovirus II type antigen and at least one porcine pseudorabies virus antigen. The invention also relates to two preparation methods of the double-combination vaccine. One method comprises the steps: inactivating the porcine circovirus II type to obtain a porcine circovirus II type inactivated vaccine; and mixing the porcine pseudorabies virus antigen with a freeze-drying protective agent to obtain a porcine pseudorabies freeze-drying living vaccine, and then dissolving the porcine pseudorabies freeze-drying living vaccine with the inactivated vaccine as a diluent to obtain the porcine circovirus II type-porcine pseudorabies double-combination vaccine composition. The other method comprises the steps: mixing the porcine circovirus II type antigen with the porcine pseudorabies virus antigen in proportion, and then mixing with the freeze-drying protective agent to obtain the double-combination freeze-drying vaccine. With use of the double-combination vaccine, the cold chain operating cost can be greatly reduced; and the combination vaccine has a synergistic role in the immune effect on the porcine pseudorabies virus antigen, and does not affect the immune effect of the porcine circovirus antigen.

Description

technical field [0001] The invention belongs to the technical field of vaccines, and in particular relates to a dual vaccine of porcine circovirus type Ⅱ and porcine pseudorabies and its preparation method and application. Background technique [0002] Porcine circovirus (PCV) is the smallest animal virus ever discovered. PCV is known to have two serotypes, PCV1 and PCV2. PCV1 is a non-pathogenic virus, and PCV2 is a pathogenic virus. In contrast to PCV1, which is usually not virulent, PCV2 infection in pigs has been associated with a variety of disease syndromes, collectively known as porcine circles. Circovirus disease, also known as porcine circovirus-associated disease. [0003] Porcine circovirus disease affects pigs 5-22 weeks of age and is clinically characterized by emaciation, pallor, growth retardation, respiratory distress, diarrhea, jaundice, and jaundice. In some affected pigs, a combination of all symptoms is evident, while others have only one or two of the...

Claims

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Application Information

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IPC IPC(8): A61K39/245A61K39/12A61P31/20A61P31/22
Inventor 张许科孙进忠白朝勇
Owner PU LIKE BIO ENG
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