Chicken C type avian metapneumovirus strain aMPV/C-JCZ and application thereof

An avian metapneumovirus, C-JCZ technology, applied in the direction of viruses/phages, antiviral agents, medical preparations containing active ingredients, etc., can solve the problems of poultry growth performance and economic impact.

Active Publication Date: 2016-02-03
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since there is no vaccine against avian metapneumovirus infection in China at present, avian metapneumovirus infection has a serious ...

Method used

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  • Chicken C type avian metapneumovirus strain aMPV/C-JCZ and application thereof
  • Chicken C type avian metapneumovirus strain aMPV/C-JCZ and application thereof
  • Chicken C type avian metapneumovirus strain aMPV/C-JCZ and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1a

[0021] Isolation and identification of embodiment 1aMPV / C-JCZ virus

[0022] 1. Collection of disease materials and isolation of viruses:

[0023] The diseased material came from an affected chicken farm in southeastern my country, and the affected chickens had severe respiratory clinical symptoms, which seriously affected the growth performance of the chickens.

[0024] Aseptically collect the turbinates, trachea, larynx and lungs of the diseased chickens, weigh them and add sterilized PBS containing 1% penicillin and streptomycin in a weight ratio of 1:4, after grinding, centrifuge at 3500rpm for 15 minutes, and The supernatant was stored in a -80°C freezer.

[0025] 2. Animal regression experiment

[0026] Randomly group 2-week-old SPF chickens, inoculate 200 microliters of tissue-derived virus with nasal drops and eye drops, and observe the clinical symptoms of the experimental group every day. On the second day after the inoculation, individual chickens began to have we...

Embodiment 2

[0030] Embodiment 2SPF chick embryo measures virus titer

[0031] Preserved virus can be directly inoculated with SPF chicken embryos through the allantoic cavity to detect the virus titer. First, the preserved virus was serially diluted 10 times, and 5 SPF chicken embryos were inoculated respectively, with 200 microliters per embryo. Incubate in an incubator at 38° C. with a humidity of 40%, observe for 24 hours and discard dead embryos, and then observe the death of chicken embryos every 8 hours. One week after inoculation, place the chicken embryos in a refrigerator at 4°C to cool. After disinfection, the tweezers break the eggshell, remove the shell membrane, tear the chorioallantoic membrane, and suck out the allantoic fluid, which contains a large amount of virus. After collection, freeze it at -80°C for storage ; Take out the chicken embryos at the same time, and observe the death or weakening of the chicken embryos (chicken embryos stop developing, bleeding or hair th...

Embodiment 3a

[0032] The establishment of embodiment 3aMPV / C challenged virus model and the determination of minimum challenged poison dose

[0033]Randomly divide 2-week-old SPF chickens, 10 chickens in each group, inoculate 100EID50, 200EID50, 500EID50, 1000EID50, 5000EID50, 10000EID50 of aMPV / C-JCZ by intranasal and eye drops, observe the clinical symptoms of the experimental group every day, and the chickens in the 100EID50 inoculation group No disease was found, and only 2 in the 200EID50 group showed mild respiratory symptoms. In the 500EID50 inoculation group, 4 chickens had obvious respiratory symptoms on the 3rd day after inoculation. All chickens fell ill on the 5th day (see Table 1). In the process of vaccine development, it is necessary to establish an experimental animal challenge model. Chicken type C avian metapneumovirus is a new infectious disease. At present, there is no designated challenge model in China to evaluate the vaccine effect. The clinical incidence of SPF chi...

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Abstract

The invention provides a chicken C type avian metapneumovirus strain aMPV/C-JCZ and application thereof. The preservation number of the virus strain is CGMCC No.10889. The virus is a chicken-source avian metapneumovirus C subtype wild virus strain. When used for conducting chicken regression experiments on SPF chickens, the wild-isolated virus causes serious respiratory tract infection clinical symptoms and leads to trachea and lung pathological injuries, and M gene sequencing verifies that the virus is C subtype avian metapneumovirus. The titer of the virus is determined on SPF chicken embryos after the virus is diluted, and the experiment result shows that EID 50 is 10<-4.6>/0.2 ml. It is the first time to isolate the chicken C type avian metapneumovirus at home, and the chicken C type avian metapneumovirus has large difference from A subtype or B subtype avian metapneumovirus in genotype and antigen. Direct supports are provided for further studying the pathogenic mechanism of aMPV and developing study on vaccine for effectively preventing and controlling the virus.

Description

technical field [0001] The invention belongs to the field of microbial animal viruses, and in particular relates to a chicken C-type avian metapneumovirus strain and related applications thereof. Background technique [0002] Avian Metapneumovirus (aMPV) belongs to the Metapneumovirus genus of the Paramyxoviridae family, and is an enveloped, non-segmented, single-stranded negative-sense RNA virus. Avian metapneumovirus infection is an emerging infectious disease that causes acute, highly contagious upper respiratory tract infection and decreased egg production in poultry such as turkeys and chickens. Avian metapneumovirus infection is worldwide. The virus is divided into 4 subtypes: A, B, C, and D. Type A or B is isolated from most parts of the world such as Europe, Asia, and South America. Type C infections have been reported in the United States, France, and South Korea. Type D infection is currently only found in France. Nucleotide sequence analysis shows that the type...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/155A61P31/14C12R1/93
Inventor 刘爵韦莉阎旭王菁朱珊珊全荣侯磊李子璇
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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