Cell culture method of chlamydia abortus and application

A technology of chlamydia abortus and its culture method, which is applied in the biological field, can solve the problems of cumbersome steps, complex production process, and inapplicability to large-scale production of antigens, and achieve the effects of portable operation, improved culture efficiency, and high-efficiency infection

Active Publication Date: 2020-09-15
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This culture method, although effective, is cumbersome and not suitable for large-scale production of antigens
Although chicken embryos can be used to produce antigens on a large scale, the production process is too complicated, and chicken embryos are considered living organisms, which has certain ethical controversies and does not conform to the development concept and trend of modern vaccine production technology

Method used

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  • Cell culture method of chlamydia abortus and application
  • Cell culture method of chlamydia abortus and application
  • Cell culture method of chlamydia abortus and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1 is based on McCoy cell culture Chlamydia abortus

[0035] 1 Material method

[0036] 1.1 Strains and cell lines

[0037] Chlamydia abortus SX5 strain, isolated from a sample of aborted dairy cows, is a vaccine-producing strain for Chlamydia inactivated vaccines in dairy cows, and has been approved as a national Class 1 new veterinary drug certificate. Chlamydia abortus GN6 strain is a clinical isolate isolated from aborted fetuses of yaks. The two strains of Chlamydia abortus were deposited in Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences. The Chlamydia abortus strain was subcultured in McCoy cells, and the titer was measured after harvesting, and stored as seeds at -80°C for future use. The titer of the Chlamydia abortus strain is expressed in inclusion body forming units per milliliter (IFU / mL), and the inoculation amount on the cells is expressed in the multiplicity of infection (MOI), which is the IFU of Chlamydia abor...

Embodiment 2

[0055] Example 2 Large-scale proliferation of Chlamydia abortus in spinner bottles based on BHK-21 cells

[0056] 1 Material method

[0057] 1.1 Strains and cell lines

[0058] The SX5 strain of Chlamydia abortus is a seedling strain for dairy cow chlamydia inactivated vaccine, which is preserved in Lanzhou Veterinary Research Institute of Chinese Academy of Agricultural Sciences. Chlamydia abortus SX5 strain was subcultured in McCoy cells, the titer was measured after harvesting, and stored as seeds at -80°C for future use. The titer of Chlamydia abortus is expressed in inclusion body forming units per milliliter (IFU / mL), and the inoculum amount on the cells is expressed in multiplicity of infection (MOI), which is the ratio of the IFU of Chlamydia abortus in the inoculum to the number of infected cells , that is, every IFU of Chlamydia abortus infects 1 cell.

[0059] Small hamster kidney cells BHK-21 were purchased from the Cell Bank of the Type Culture Collection Commi...

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Abstract

The invention relates to a cell culture method of chlamydia abortus and application. The culture method disclosed by the invention comprises the following steps: washing cells growing into a monolayerto obtain washed cells; mixing the chlamydia abortus inoculation liquid with a DNA transfection reagent Lipofectamine 2000, allowing the mixture to stand, inoculating the washed cells with the mixture, incubating, sucking and discarding the incubated liquid, and culturing for 48-72 hours by using a culture solution. According to the culture method, centrifugal operation is not needed, efficient infection of chlamydia abortus on in-vitro culture cells can be achieved, and operation is convenient.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a cell culture method and application of chlamydia abortus. Background technique [0002] Chlamydia spp. is a Gram-negative, obligate intracellular parasitic prokaryotic microorganism with a unique biphasic developmental cycle. It can infect humans and various animals and cause diseases. It is an important class of pathogenic microorganisms. All Chlamydia species have the same life cycle, including 2 stages of infection and intracellular proliferation. The primary bodies of Chlamydia (elementary bodies, EBs) are metabolically inactive but infectious. After the free EBs attach to the surface of host cells, they enter the cells through endocytosis and form inclusion bodies (inclusions). In the inclusion bodies, EBs transform into reticulate bodies (RBs) with vigorous metabolism, and RBs proliferate by binary fission. After a certain period of proliferation, RBs re-transformed into EB...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N5/077C12N5/071C12R1/01
CPCC12N1/20C12N5/0656C12N5/0686
Inventor 周继章李兆才刘萍刘东慧
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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