Vaccine strain of HIN1 swine influenza virus and application thereof

A technology of swine influenza virus and influenza A virus, applied in the field of animal virology, can solve the problems of short duration of antibody, low level of antibody, and unsatisfactory immunogenicity and reactogenicity, and achieve long duration of antibody , good immunogenicity and reactogenicity, the effect of preventing H1N1 swine flu

Active Publication Date: 2013-01-30
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The immunogenicity and reactogenicity of swine influenza virus vaccine strains in the prior art cannot meet the requirements, and the antibody level produced by the vaccine prepared by adopting the existing vaccine strains is not high, and the antibody duration is not long, and cannot effectively resist the current epidemic virus. strain challenge

Method used

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  • Vaccine strain of HIN1 swine influenza virus and application thereof
  • Vaccine strain of HIN1 swine influenza virus and application thereof
  • Vaccine strain of HIN1 swine influenza virus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Purification and Identification of Porcine Influenza A Virus A / Swine / Nanjing / 50 / 2011(H1N1) Strain

[0025] 1. Virus Isolation and Harvest of Chicken Embryo Allantoic Fluid

[0026] Take the lung tissue of the infected pig and add an appropriate amount of sterilized saline to grind it into a homogenate, freeze and thaw three times, centrifuge at 8000 rpm for 10 minutes, discard the upper layer of fat, and absorb the middle liquid. Add penicillin at a final concentration of 1000 units / ml and streptomycin at a final concentration of 1000 units / ml to the intermediate liquid and treat at 4°C for 1 hour, then centrifuge at 8000 rpm for 10 minutes, and take the supernatant. The supernatant was inoculated into 9-11-day-old SPF chicken embryos through the allantoic cavity, 0.2ml / piece, and cultured in an incubator for 72 hours. Observe twice a day, remove the contaminated embryos within 24 hours, and collect the allantoic fluid of the remaining chicken embryos.

[...

Embodiment 2

[0043] Example 2 Determination of the whole genome sequence of purified virus

[0044] 1. Synthesis of RT-PCR primers

[0045] Referring to the published SIV sequence data on Genbank, we designed and synthesized random primers Unil2 and RT-PCR primers for 8 gene segments (PB2, PB1, PA, HA, NA, NP, NS, M) of the SIV genome (see Table 2 ), were synthesized by Nanjing GenScript Biotechnology Co., Ltd. Wherein the upstream primer PB2-F and the downstream primer PB2-R are used to amplify the PB2 fragment; the upstream primer PB1-F and the downstream primer PB1-R are used to amplify the PB1 fragment; the upstream primer PA-F and the downstream primer PA-R are used for Amplify PA-R fragment; upstream primer NP-F and downstream primer NP-R are used to amplify NP fragment; upstream primer NS-F and downstream primer NS-R are used to amplify NS fragment; upstream primer M-F and downstream primer M-R It is used to amplify the M fragment; the upstream primer HA-F and the downstream pri...

Embodiment 3

[0063] Example 3 Preparation of Inactivated H1N1 Swine Influenza Virus Vaccine and Its Protective Efficacy Experiment

[0064] 1. Materials and methods

[0065] (1) Main reagents

[0066] Esso white oil Marcol 52 was purchased from Nanjing Tianbang Biotechnology Co., Ltd., batch number: vg4552975P.

[0067] Tween-80 CRILLET 4, purchased from Singapore CRODA company, batch number: 15448

[0068] Span-80 CRILL 4, purchased from Singapore CRODA company, lot number: 15438

[0069] (2) Test pigs

[0070] Ten H1N1 subtype swine influenza antibody-negative (HI) healthy 30-40-day-old weaned piglets were provided by a pig farm in Hongze, Jiangsu.

[0071] (3) Antigen preparation

[0072] Dilute chicken embryo allantoic fluid containing H1N1 subtype swine influenza virus A / Swine / Nanjing / 50 / 2011 strain 1000 times, then inoculate 9-11-day-old SPF or non-vaccinated chicken embryos at 0.1ml / piece, and discard for 24 hours Chicken embryos that died within 72-96 hours were placed at 4...

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Abstract

The invention relates to a vaccine strain of HIN1 swine influenza virus, wherein a classification naming is Swine influenza virus A / Swine / Nanjing / 50 / 2011 (H1N1), and a microbial accession number is CCTCCNO: V201218. According to the invention, isolation, identification and whole genome sequencing of the HIN1 swine influenza virus strain are carried out, and an inactivated vaccine against the swine flu is prepared by using the HIN1 swine influenza virus strain. The inactivated vaccine against the H1NI swine flu prepared from the virus strain can provide good protection for pigs attacked by homologous virulent strains, has good immunogenicity, and can effectively prevent H1NI swine flu with single vaccine or combined vaccines.

Description

technical field [0001] The invention relates to the field of animal virology, in particular to an H1N1 type swine influenza virus vaccine strain and its application in preparing medicines for preventing or treating swine influenza. Background technique [0002] Swine influenza virus (SIV) belongs to Orthomyxoviridae type A influenza virus, which can cause acute, febrile, mass and high contact respiratory diseases in pigs, manifested as sudden high fever, depression, loss of appetite, Difficulty breathing, paroxysmal cough and other symptoms. Although the mortality rate of a single swine flu disease is not high, and sick pigs can recover quickly, it is often co-infected with other diseases such as blue ear disease, infectious pleuropneumonia, and streptococcus suis, which complicates the condition. Serious, resulting in a sharp rise in pig mortality, resulting in serious economic losses. In addition, the pig body can act as a "mixer" for human influenza and avian influenz...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/145A61P31/16C12R1/93
Inventor 白昀冯志新邵国青刘茂军白方方熊祺琰王海燕王占伟甘源刘冬霞
Owner JIANGSU ACAD OF AGRI SCI
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