Caprine parainfluenza virus type 3 JS14-2 strain and application thereof

A parainfluenza and virus technology, which is applied to the goat parainfluenza virus type 3 JS14-2 strain and its application field to achieve the effects of preventing epidemics, good immunogenicity and stable biological characteristics

Active Publication Date: 2018-12-07
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Vaccines and diagnostics to prevent goat parainfluenza virus type 3 infection are not yet available

Method used

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  • Caprine parainfluenza virus type 3 JS14-2 strain and application thereof
  • Caprine parainfluenza virus type 3 JS14-2 strain and application thereof
  • Caprine parainfluenza virus type 3 JS14-2 strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Isolation, purification and identification of goat parainfluenza virus type 3 JS14-2 strain

[0032] 1 Virus isolation

[0033] In 2013, many large-scale goat farms in Jiangsu, Anhui and other places in my country had diseases with respiratory symptoms as the main manifestations, manifested as depression, cough, serous fluid or thick nasal fluid. The nasal swab samples collected from the sick goats were detected and identified as goat parainfluenza virus type 3. Subsequently, we continued to carry out etiological testing on many similar cases in Jiangsu, and collected nasal swab samples from the affected sheep farms and stored them at -80°C.

[0034] Add penicillin and streptomycin to the nasal swab samples of infected goats detected as positive for parainfluenza virus, incubate at 37°C for 30min, centrifuge at 12000r / m for 20min, take the supernatant and filter it through a 0.22μm filter membrane, and inoculate it into bovine kidney cells ( MDBK) (purchased...

Embodiment 2

[0040] Example 2 Hemagglutination analysis

[0041] The hemagglutination value of the isolated and purified goat parainfluenza virus type 3 was determined by hemagglutination test. Different virus solutions were diluted 2-fold on a 96-well hemagglutination plate, then added with 1% guinea pig red blood cells, and incubated at 37°C for 45 minutes, and the highest dilution that could completely agglutinate the red blood cells was used as the hemagglutination value of the virus. The results showed that the hemagglutination value could reach 256 as measured by the hemagglutination test.

Embodiment 3

[0042] Example 3 Morphological Observation of Virus Particles

[0043] Take goat parainfluenza virus type 3 JS14-2 strain cell culture 200mL, centrifuge at 12000r / min for 5min to remove cell debris, supernatant is ultracentrifuged at 40000r / min for 2h, take the precipitate and dissolve it in PBS overnight to obtain virus suspension. The virus suspension was loaded on the copper grid, and after negative staining with 2% phosphotungstic acid, the morphology of virus particles was observed under a transmission electron microscope. Such as figure 1 As shown, a large number of enveloped virus particles can be seen, and the virus presents an irregular shape with a size of about 100-150nm.

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Abstract

The invention belongs to the field of veterinary biological products and discloses a caprine parainfluenza virus type 3 JS14-2 strain and an application thereof in preparation of inactivated vaccines.The inactivated vaccines are prepared and obtained by the steps of: separating and purifying to obtain the caprine parainfluenza virus type 3 JS14-2 strain, carrying out centrifugation and formaldehyde inactivation on virus liquid and adding an adjuvant for emulsification. The invention provides a virus strain which can be used for preparing caprine parainfluenza virus type 3 inactivated vaccines. The strain belongs to domestic prevalent strains, and is strong in virulence and good in antigenicity; the strain has better growth characteristic on MDBK (Madin-Darby Bovine Kidney) cells, and canobtain stable high titer (more than or equal to 107 TCID50 / mL, and the hemagglutination valence is more than or equal to 28); the vaccine preparation method is perfect, and the immunogenicity and theimmune protective effect are good; the used immunizing dosage is low, the safety is high, the immune duration is long, the production and use cost is greatly saved, and the benefit for effectively controlling the caprine parainfluenza virus type 3 in clinical practices is achieved.

Description

technical field [0001] The invention belongs to the field of veterinary biological products, and relates to a goat parainfluenza virus type 3 JS14-2 strain and application thereof. Background technique [0002] Caprine parainfluenza virus 3 (CPIV3) is a newly identified member of the Respirovirus genus in the Paramyxoviridae family. It is an enveloped RNA virus that mainly infects goats and sheep. The virus is mainly transmitted through the respiratory tract and causes respiratory diseases. Under the conditions of stress, mixed or secondary infection with other pathogens, it will cause obvious clinical symptoms, cause severe respiratory diseases, and lead to high morbidity and mortality. Epidemiological studies have shown that there is a high virus infection rate in domestic sheep flocks. The virus is a newly discovered new pathogen, and there are no reports on effective prevention and control methods for the virus at home and abroad. There are no vaccines or diagnostic re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12Q1/70C12Q1/686C12R1/93
CPCC12N7/00C12N2760/18621C12N2760/18651C12Q1/686C12Q1/701C12Q2521/107
Inventor 李文良毛立郝飞李基棕杨蕾蕾张纹纹王钟毓孙敏刘茂军江杰元
Owner JIANGSU ACAD OF AGRI SCI
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