Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A kind of culture method of porcine circovirus and its application

A technology of porcine circovirus and a culture method, applied in the field of veterinary biological products, can solve the problems of easy invasion, incompatibility with production, weakened immunity of the respiratory tract, etc.

Active Publication Date: 2018-02-09
PU LIKE BIO ENG
View PDF17 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The Ross study at the University of Iowa found that after Mhp infection, the ability of lymphocytes to produce antibodies decreased, cellular immunity decreased, and the ability of alveolar macrophages to phagocytize and clear pathogens also decreased, and the activity of suppressive T cells increased, resulting in respiratory immunity. Weakened disease resistance, which makes it easier for other pathogens to invade, and can cause secondary infection of porcine reproductive and respiratory syndrome, porcine circular ring, etc., resulting in the failure of various vaccinations
[0006] The biological characteristics of PCV2 are quite special. Although it can exist in the body at a high titer for a long time, its proliferation titer is low and unstable in vitro culture. It is difficult to exceed the highest level of PCV2 in the international application of PK15 subcultured cells. 10 3.5 TCID 50 / ml; and the results of immunofluorescence test show that the proportion of PK15 cells infected by PCV2 is only 20%-30%, so the titer of PCV2 toxin production is low, which is not suitable for the current large-scale animal vaccine production (Shen Lu. Zhu Yuan The Establishment of Fluorescent Quantitative PCR Detection Method for Cyclovirus Type 2 and Preliminary Discussion on the Culture Technology. Master's Thesis of Nanjing Agricultural University)

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A kind of culture method of porcine circovirus and its application
  • A kind of culture method of porcine circovirus and its application
  • A kind of culture method of porcine circovirus and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] The cultivation of different components of embodiment 1 and the mensuration of content

[0074] 1.1 Cultivation of different components

[0075] 1.1.1 PCV2 culture alone

[0076] To cultivate PK15 cells, drain the cell culture medium from the monolayer PK15 cells that have grown to 80%-100%.

[0077] Inoculate the basic seed virus of PCV2 SH strain on PK15 cells at 0.01 MOI, absorb at 37°C for 30 minutes, add the cells prepared by MEM medium containing 4% (V / V) calf serum and 2mmol / l D-glucosamine hydrochloride to maintain cultured at 37°C for 5-7 days, freeze-thawed at -40°C for 2-3 times, and the virus was harvested as culture 1.

[0078] 1.1.2 Co-culture of PCV2 and Mhp

[0079] To cultivate PK15 cells, drain the cell culture medium from the monolayer PK15 cells that have grown to 80%-100%.

[0080] Inoculate PCV2 SH strain and Mycoplasma hyopneumoniae HN0613 strain basic seed virus / bacteria into PK15 cells at 0.01 MOI of PCV2SH strain and 1% (V / V) of Mycoplasma ...

Embodiment 2

[0094] Co-culture of embodiment 2 PCV2 and Mhp

[0095] 2.1 Co-culture of PCV2 and Mhp

[0096] Prepare the co-culture of PCV2 and Mhp according to Example 1.1.2, wherein the PCV2 strains are respectively SD strain and WH strain, that is, respectively cultivate the co-culture of PCV2 SD strain and Mycoplasma hyopneumoniae HN0613 strain, and PCV2 WH strain and pig The co-culture of Mycoplasma pneumoniae HN0613 strain was named as culture 5 and culture 6 in turn.

[0097] 2.2 Determination of PCV2 and Mhp content in culture 5 and culture 6

[0098] According to the method in Example 1.2, the contents of PCV2 and Mhp in culture 5 and culture 6 were measured respectively, and the measurement results are shown in Table 2.

[0099] Table 2 Determination results of different components after cultivation

[0100]

Embodiment 3

[0101] Example 3 Preparation of PCV2 Single Vaccine and PCV2-Mhp Vaccine Composition

[0102] 3.1 Inactivation and inspection

[0103] Culture 1, culture 2, culture 5 and culture 6 prepared in Example 1-2 were inactivated by directly adding 10% formaldehyde solution at 0.1% (V / V), inactivated at 37°C for 24 hours, each Stir every 4 hours. The inactivated culture 1 and culture 2 were sampled for sterility and inactivation test respectively, and the culture 1, culture 2, culture 5 and culture 6 that passed the inspection were respectively used as antigen solutions and stored at 4°C.

[0104] 3.2 Preparation of Mycoplasma hyopneumoniae single vaccine

[0105] Prepare according to the method of Mycoplasma hyopneumoniae described in Chinese patent CN103127497A (Zhang Xuke, Sun Jinzhong, Bai Chaoyong. Porcine circovirus type 2, Mycoplasma pneumoniae dual inactivated vaccine and preparation method thereof. Chinese patent CN103127497A), and obtain Mycoplasma hyopneumoniae concentrat...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
diameteraaaaaaaaaa
Login to View More

Abstract

The invention provides a method for cultivating porcine circovirus and its application, and a method for cultivating mycoplasma hyopneumoniae. The method for cultivating porcine circovirus can simultaneously cultivate porcine circovirus and mycoplasma hyopneumoniae, not only can multiply the titers of porcine circovirus and mycoplasma hyopneumoniae simultaneously, but also can obtain high titer porcine circovirus. The vaccine composition prepared by using the porcine circovirus antigen prepared by the cultivation method and the mycoplasma hyopneumoniae antigen can prevent and / or treat diseases caused by the porcine circovirus and mycoplasma hyopneumoniae.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a method for cultivating porcine circovirus and its application, and a method for cultivating mycoplasma hyopneumoniae. Background technique [0002] In 1974, German scholar Tischer et al. detected a cell pollutant from the continuously passaged pig kidney cell line PK15 cells, which was a virus-like particle that could not cause cytopathic changes. In 1982, the scholar further research confirmed that the genome of the virus is composed of single-stranded circular DNA, thus naming it Porcine Circovirus (PCV). [0003] Porcine circovirus belongs to the genus Circovirus of the family Circoviridae and is the smallest animal virus ever discovered, with an average diameter of 17 nm. According to the difference of pathogenicity, antigenicity and nucleotide sequence of porcine circovirus, PCV is divided into two genotypes of non-pathogenic PCV1 and pa...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00C12N1/20A61K39/12A61P31/20C12R1/35C12R1/93
Inventor 刘守川宋国亮彭伍平张许科孙进忠王延辉王莹田克恭
Owner PU LIKE BIO ENG
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com