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Virus velogen strain for porcine reproductive and respiratory syndrome, attenuated vaccine strain thereof and application thereof

A technology for respiratory syndrome and attenuated vaccine, which is applied in the invention also relates to the application of attenuated vaccine strains in the prevention or treatment of highly pathogenic porcine blue-ear disease, and achieves the effect of good safety.

Active Publication Date: 2008-10-08
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can be seen that although there are many kinds of PRRS vaccines at present, none of them can produce enough protective effect on pigs

Method used

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  • Virus velogen strain for porcine reproductive and respiratory syndrome, attenuated vaccine strain thereof and application thereof
  • Virus velogen strain for porcine reproductive and respiratory syndrome, attenuated vaccine strain thereof and application thereof
  • Virus velogen strain for porcine reproductive and respiratory syndrome, attenuated vaccine strain thereof and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Isolation and identification of porcine reproductive and respiratory syndrome virus virulent strain HuN4

[0032] 1. Materials and methods

[0033] 1.1Marc-145 cells, monoclonal antibodies against PRRSV-N and GP5 protein are preserved by Harbin Veterinary Research Institute.

[0034] 1.2 Collection and processing of disease materials Anticoagulant blood, lungs, kidneys and lymph nodes of pigs with high fever in a pig farm in a southern city were collected.

[0035] 1.3 Marc-145 cell culture Take out 2-3 ampoules of frozen Marc-145 cells from liquid nitrogen, and thaw quickly in a 37°C water bath. Centrifuge each ampoule at 1000r / min for 5-10min, discard the supernatant, and use 2-3ml of DMEM containing 100IU / ml penicillin, 80μg / ml streptomycin, 6% calf serum and 0.5% hydrolyzed milk protein Resuspend the cells, then inhale into a culture flask containing 10ml of culture medium, shake well, and store at 37°C, 5% CO 2 After 72 hours, the culture was expanded ...

Embodiment 2

[0070] Example 2 Breeding of Porcine Reproductive and Respiratory Syndrome Virus Attenuated Vaccine Strain HuN4-F112

[0071] 1. Material method

[0072] 1.1 Virus cell culture and cloning Marc-145 cells for virus cultivation were cultured with DMEM cell culture medium containing 8% inactivated calf serum, 100U / ml penicillin, 80μg / ml streptomycin, and NaHCO 3 Adjust the pH value to about 7.2, and filter to sterilize. Use this cell culture medium to resuscitate Marc-145 cells or pass them to a sterilized cell culture flask with a capacity of 100ml, and use it to inoculate the virus after growing into a good cell monolayer.

[0073] The cloning of 1.2 kinds of viruses The highly pathogenic porcine PRRS virus HuN4 strain that embodiment 1 separates is passed on Marc-145 cell, carry out virus plaque clone when passing on to 3,15,20 generations respectively, and Plaques with a diameter of 1 mm were selected to continue subculture.

[0074] 1.3 Determination of the toxicity of th...

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Abstract

The invention discloses a virulent strain HuN4 for porcine reproductive and respiratory syndrome, and an attenuated vaccine strain HuN4-F112 (the collection number of microorganism: CGMCC No.2484) attenuated because of virulent strain HuN4 passage, and also discloses the application of the attenuated vaccine strain in preventing or curing the highly pathogenic blue-ear disease, which belongs to the biomedical field. The attenuated vaccine strain HuN4-F112 can be prepared into single-vaccine or combined strain (live vaccine or inactivated vaccine), can effectively prevent or cure the highly pathogenic blue-ear disease, and also can be prepared into diagnostic reagent for the highly pathogenic blue-ear disease diagnosis. The attenuated vaccine strain HuN4-F112 of the invention has the advantages of good security and high protection efficiency.

Description

technical field [0001] The invention relates to a virulent strain of virus and an attenuated vaccine strain attenuated by it, in particular to a virulent strain of porcine reproductive and respiratory syndrome virus HuN4, and an attenuated vaccine strain HuN4 attenuated by the virulent strain HuN4 - F112, the present invention also relates to the application of the attenuated vaccine strain in the prevention or treatment of highly pathogenic pig PRRS, which belongs to the field of biomedicine. Background technique [0002] In the spring and summer of 2006, pig "high fever disease" characterized by fever, skin redness, high morbidity and high mortality occurred in some parts of southern my country, and then gradually spread to most provinces in the country. A large number of pathogens were isolated and identified, and animal experiments confirmed that the main pathogen of pig "high fever" is a mutant strain of highly pathogenic porcine blue-ear disease virus, which is also kn...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12N7/08A61K39/12A61P31/14C12Q1/70
Inventor 童光志田志军周艳君安同庆彭金美
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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