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Raccoon canine distemper virus attenuated vaccine strain and its application in preparing live raccoon distemper vaccine

A technology of attenuated vaccine and raccoon dog distemper, which is applied in the direction of virus, antiviral agent, virus/phage, etc., can solve the problems of lack of special treatment methods and inability to obtain better immune protection effect, and achieve good immunogenicity, drop degree of stability

Active Publication Date: 2020-01-14
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because raccoon distemper has the characteristics of acute onset and lack of effective treatment methods, vaccination is still the main prevention and control method for raccoon distemper. However, there is no raccoon distemper vaccine on the market at present. The canine distemper vaccine of other animals is used to prevent raccoon canine distemper, but it cannot achieve better immune protection effect

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Embodiment 1 Domestication and Identification of Canine Distemper Virus Vaccine Strain CDV-O2

[0012] 1. Domestication of canine distemper virus vaccine strain CDV-O2

[0013] (1) Inoculate the preserved canine distemper virus vaccine strain CDV-O2 on African green monkey kidney cells (Vero) by adsorption and inoculation to carry out virus amplification and culture, observe the cell virus situation every day, and wait for the cytopathic changes to reach 80 % or so, the virus liquid was harvested by the method of repeated freezing and thawing, and the virus content (TCID 50 ≥10 5.0 / ml), sterility test. Aliquot the tested virus liquid and store it at -20°C.

[0014] (2) The amplified CDV-O2 virus liquid stock solution was inoculated into the hindlimb muscles of 3 antibody-negative healthy raccoon dogs, 5ml each, inoculated at three points, kept in isolation, and observed the physiological indicators of the inoculated raccoon dogs every day. On the 3rd and 5th day af...

Embodiment 2

[0021] Preparation of embodiment 2 raccoon dog distemper live vaccine (CDV-OR12 strain)

[0022] 1. Preparation of Canine Distemper Virus Seedling Solution

[0023] Take the preserved canine distemper virus attenuated vaccine strain CDV-OR12 and inoculate it on Vero cells by adsorption and inoculation for virus amplification and culture. Observe the cell virus situation every day. Methods The virus liquid was harvested and tested for virus content and purity. The content of raccoon canine distemper virus attenuated vaccine strain in the obtained virus liquid was TCID 50 ≥10 4.5 / ml. According to the measured virus content data, after appropriate dilution of the tested virus liquid with serum-free MEM, quantitatively dispense into 90ml vaccine bottles and store at -20°C.

[0024] 2. Determination of canine distemper virus content in finished vaccine products

[0025] Vaccine finished product is tested for virus content, canine distemper virus content TCID 50 for 10 4.5 -1...

Embodiment 3

[0026] Safety test of embodiment 3 raccoon dog distemper live vaccine (CDV-OR12 strain)

[0027] Get the qualified raccoon dog distemper live vaccine (prepared in embodiment 2), hindlimb muscle inoculation antibody negative raccoon dog, establish single dose (vaccinate 1 part, canine distemper virus content TCID 50 =10 4.5 / head), single-dose repeated and super-dose (10 times) immunization groups, and non-vaccinated raccoon dogs were set as negative controls. After vaccination, the body temperature, appetite, mental state, feces and other physiological indicators of raccoon dogs were observed for 14 consecutive days, and the lesions of the main organs of vaccinated raccoon dogs were observed by autopsy. The test results showed that after the single-dose, repeated single-dose and over-dose inoculation, all the physiological indexes of the raccoon dog were normal and did not show any clinical symptoms. Safe for raccoons.

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Abstract

The invention discloses an attenuated vaccine strain of raccoon canine distemper virus and its application in preparing a live vaccine of raccoon canine distemper, which belongs to the technical field of veterinary biological products. The present invention artificially domesticates the canine distemper vaccine strain CDV‑O2 through the continuous passage of raccoon dogs, and obtains an attenuated canine distemper virus vaccine strain that is safe to raccoon dogs, has good immunogenicity and stable titer, and is named CDV ‑OR12 strain, the vaccine strain is preserved in the General Microbiology Center of China Microbiology Culture Collection Management Committee, and its microorganism preservation number is CGMCC NO.13793. The live vaccine can effectively protect raccoon dogs from the attack of canine distemper virus, the protection rate is above 90%, and can effectively prevent the occurrence of raccoon dog distemper, and the immunity period is 6 months. The proposal of the present invention provides a new technical means for the prevention and control of raccoon dog distemper.

Description

technical field [0001] The invention relates to an attenuated vaccine strain of raccoon canine distemper virus and its application in the preparation of live raccoon distemper vaccine, belonging to the technical field of veterinary biological products. Background technique [0002] Canine distemper is an acute, febrile, and highly contagious infectious disease caused by canine distemper virus of the genus Morbillivirus in the Paramyxoviridae family. Most of the young animals have an acute and fatal course, and adult animals can become chronic and persistent. Infection, the clinical symptoms are mainly characterized by biphasic elevated body temperature, accompanied by catarrhal pneumonia and gastroenteritis, and occasionally neurological symptoms appear, which is prone to secondary mixed infection and secondary infection of other bacteria and viruses, and the mortality rate can be as high as 80% %, is one of the most serious diseases that endanger my country's fur animal breed...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/04A61K39/165A61P31/14C12R1/93
CPCA61K39/12C12N7/00C12N2760/18421C12N2760/18434
Inventor 闫喜军胡博吕爽王洋廉士珍刘昊张蕾白雪鲁荣光张海玲赵建军朱言柱史宁薛向红邓效宇徐淑娟
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS