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Infectious bronchitis low-virulent live vaccine YX10 D90 strain

A technology for bronchitis and chicken infectivity, applied in the direction of viruses, antiviral agents, virus antigen components, etc., can solve the problems of ineffective prevention and control of chicken infectious bronchitis virus epidemic strains, unsatisfactory control effects, etc. , to achieve the effect of wide application value

Inactive Publication Date: 2016-06-15
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome the defect that the existing vaccine strains cannot effectively prevent the attack of the popular strain of chicken infectious bronchitis virus, and the control effect is not ideal, and provide a live attenuated chicken infectious bronchitis vaccine YX10D90 strain

Method used

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  • Infectious bronchitis low-virulent live vaccine YX10 D90 strain
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  • Infectious bronchitis low-virulent live vaccine YX10 D90 strain

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Example 1 Isolation and Identification of Infectious Bronchitis Virus YX10 Strain

[0023] YX10 was isolated and identified by our laboratory in 2010 from a 30-day-old diseased yellow-feathered broiler chicken in Huzhou, Zhejiang. The specific process is as follows:

[0024] (1) Isolation and culture of virus

[0025] Aseptically collect the kidneys, spleen, lungs, and trachea of ​​dead chickens, sterilize the normal saline at a volume ratio of 1:5 (volume ratio), grind, freeze-thaw 3 times, centrifuge at 5000rpm for 20min, and filter the supernatant to sterilize the allantois Inoculate 10 10-day-old SPF chicken embryos in the cavity, inoculate 0.2ml per embryo, incubate the inoculated chicken embryos at 37°C, illuminate the embryos twice a day, discard dead embryos within 24 hours, and take 5 embryos within 48 hours to harvest the allantois solution, and the rest continued to incubate until 144 hours, and the pathological changes of chicken embryos were recorded. So ...

Embodiment 2

[0047] The cultivation of embodiment 2YX10D90 attenuated strain

[0048] 1. Continuous subculture of chicken embryos weakened the virus strain

[0049] Use 9-11 day-old SPF chicken embryos to carry out continuous subculture to weaken the YX10 strain, inoculate 10 chicken embryos through the allantoic cavity for each generation, 0.1ml / piece, incubate at 37°C, discard dead chicken embryos within 24 hours, 36 Harvest 5 live embryos in about 1 hour, collect the embryo fluid aseptically for the next subculture, and keep the rest until 144 hours to observe whether there are branched lesions in the chicken embryos. Such continuous passage to the 120th generation. During a certain interval, virus content, sterility and mycoplasma detection, exogenous virus detection, and safety tests on susceptible chicks were carried out to determine whether the virulence was attenuated.

[0050] 2. Determination of virus content

[0051] The EID of the D5, D25, D40, D58, D75, D90, and D120 genera...

Embodiment 3

[0071] Embodiment 3 YX10D90 attenuated vaccine preparation

[0072] 1. Preparation of IBV virus liquid

[0073]Dilute the YX10D90 species poison obtained in Example 2 1000 times with sterilized saline, inoculate 100 pieces of 10-day-old SPF chicken embryos in the allantoic cavity, 0.2ml per embryo, seal the pinholes, and continue incubation at 37°C without turning the eggs. Eggs were illuminated once within 24 hours after inoculation, and dead embryos were discarded. After that, eggs were illuminated once every 4 hours, and dead chicken embryos were taken out at any time until 36 hours. Stand upright and cool at 4°C for 12 hours. Take out the cooled chicken embryo, disinfect the air cell part, then peel off the egg shell of the air cell with aseptic operation, remove the egg shell membrane, cut the chorioallantoic membrane and amniotic membrane (do not break the yolk), and absorb the chicken embryo fluid , samples were taken for inspection, and stored at -20°C for later use....

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Abstract

The invention belongs to the field of the biotechnology and discloses an infectious bronchitis low-virulent live vaccine YX10 D90 strain. The infectious bronchitis low-virulent live vaccine YX10 D90 strain is preserved in China Center for Type Culture Collection on 14th March, 2016, the preservation number is CCTCC NO:V201610, and the preservation address is Wuhan University, Wuhan, China. The low-virulent strain is safe for all chickens and has no side reaction, the low-virulent strain is prepared into a vaccine safely and effectively, toxin attacking on homological strong toxicity can be protected, and practical and wide application value is achieved.

Description

technical field [0001] The invention relates to the field of biotechnology, and more specifically relates to a YX10D90 strain of chicken infectious bronchitis attenuated live vaccine. Background technique [0002] Infectious bronchitis (IB) is an acute, highly contagious respiratory disease of chickens caused by avian infectious bronchitis virus (IBV). The main features are respiratory symptoms in sick chickens, enlarged and pale kidneys, a large amount of urate deposits, and even the appearance of "mottled kidneys"; the egg production and egg quality of laying hens are reduced, and infection of young hens can cause permanent fallopian tubes degenerates, leading to the production of "false hens". In addition, some IBV strains can also cause lesions in the intestinal tract, glandular stomach, and muscles. IBV can infect all ages and most types of chickens, especially chicks aged 1 to 5 weeks are very susceptible, which can easily lead to growth retardation, weight gain, red...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12N15/50C07K14/165A61K39/215A61P31/14C12R1/93
CPCA61K39/12A61K2039/5254A61K2039/543C12N7/00C12N2770/20021C12N2770/20034
Inventor 谢青梅薛瑜封柯宇毕英佐
Owner SOUTH CHINA AGRI UNIV
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