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Chicken C-type acian metapneumovirus strain(aMPV-JCX) and application thereof

A technology of avian metapneumovirus and C-JCX, which is applied in the direction of virus/bacteriophage, antiviral agents, medical preparations containing active ingredients, etc., can solve the problem of decreased production performance of broiler breeders and older laying hens, and cannot fully meet the requirements of virus-induced diseases. Disease mechanism and vaccines, large losses and other issues

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

During our investigation and research, due to the infection of type C avian metapneumovirus, the production performance of broiler breeders and older laying hens dropped sharply in several enterprises, and they had to be eliminated, resulting in huge losses
Since chicken C-type avian metapneumovirus was reported for the first time in my country, we mainly isolate, cultivate and identify SPF chickens or chicken embryos, but this cannot fully meet the needs of research on the pathogenic mechanism of the virus and vaccines.

Method used

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  • Chicken C-type acian metapneumovirus strain(aMPV-JCX) and application thereof
  • Chicken C-type acian metapneumovirus strain(aMPV-JCX) and application thereof
  • Chicken C-type acian metapneumovirus strain(aMPV-JCX) and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1a

[0015] The acquisition and identification of embodiment 1a MPV / C-JCX strain

[0016] 1. Departure virus aMPV / C-JCZ

[0017] The originating virus was aMPV / C-JCZ, which was collected on June 24, 2015 in the General Microbiology Center of the China Committee for the Collection of Microbial Cultures (Address: No. 1 Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, Zip code 100101) is preserved, and the classification name is Avainmetapneumovirus (Avainmetapneumovirus), and the preservation number is CGMCCNo.10889.

[0018] Calculate the EID50 of the virus according to the Reed-Muench method, lgEID50=10 -4.6 / 0.2ml. It was confirmed by returning chickens that infection by aMPV / C-JCZ alone could cause severe respiratory symptoms in commercial yellow-feathered broiler chickens; pathological examination of the lungs and trachea revealed that virus infection caused severe inflammatory pathological damage.

[0019] Use QiagenRNeas...

Embodiment 2a

[0030] The virus titer TCID50 of embodiment 2aMPV / C-JCX strain measures

[0031] Save the virus for 10-fold serial dilution, inoculate Vero cells in a 96-well plate, at least 8 parallels for each virus dilution gradient, set up negative and positive controls, and observe the cytopathic changes in 5-7 days. Calculate the TCID50 of the virus according to the Reed-Muench method, lgTCID50=10 -4.2 / 0.1ml.

Embodiment 3a

[0032] Embodiment 3aMPV / C-JCX strain is to SPF chick embryo virulence assay

[0033] The preserved cell virus can be directly inoculated into SPF chicken embryos through the allantoic cavity to detect the virus virulence. The preserved virus was inoculated into 5 SPF chicken embryos, and each embryo was inoculated with 200 microliters. Incubate in an incubator at 38° C. with a humidity of 40%, and observe whether the chicken embryos die on time. One week after inoculation, the chicken embryos were cooled in a refrigerator at 4°C. After disinfection, the eggshells were broken with forceps, the chorioallantoic membrane was removed, and the chorioallantoic membrane was torn. At the same time, the chicken embryos were taken out to observe the development of the chicken embryos. It was found that aMPV-JCX did not cause developmental retardation or death of chicken embryos, indicating that aMPV-JCX has been stably passaged on Vero cells, and the virulence of this strain is weak, so...

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Abstract

The invention provides a chicken C-type acian metapneumovirus strain(aMPV-JCX) and application thereof. The preservation number of the aMPV / C-JCX strain is CGMCC No.10900. The virus is a cell culture adaptive strain, a preparation method of the cell culture adaptive strain comprises the steps that tissue grinding fluid of a sick chicken infected by aMPV / C-JCX which is firstly obtained through separation domestically is collected to be inoculated into single-layer Vero cells which grow being adherent to the wall, a cell culture serves as an inoculum for the next round of passage, sequentially continuous passage is conducted, and the cell culture adaptive strain is obtained. In the adaptive process, the early-generation culture cycle is 7-9 days, the culture cycle is shortened to 3-5 days when the 60th generation is cultured, and an obvious cytopathic effect can be observed. It is verified through a transmission electron microscope, indirect immunofluorescence and a western-blotting method that the aMPV / C-JCX can be effectively cultured. The titer 1 g TCID50 of the virus is equal to 10<-4.2> / 0.1 ml. It is verified through SPF chickens that the virus virulence is weak, good immunogenicity and protectiveness are achieved, and the cell-adapted virus is an ideal candidate strain for researching and producing viral vaccine.

Description

technical field [0001] The invention belongs to the field of microbial animal vaccines, and in particular relates to a chicken C-type avian metapneumovirus and its related applications. Background technique [0002] Avian metapneumovirus (aMPV) is a newly discovered and important pathogen of poultry respiratory diseases. It causes acute and highly contagious upper respiratory tract infections in poultry such as turkeys and chickens, causing egg production in breeders and laying hens. drop sharply and affect the quality of the eggshell. The virus belongs to the genus Metapneumovirus of the family Paramyxoviridae. The avian metapneumovirus observed under a negative-contrast electron microscope is polygonal and elliptical, with a common diameter of 80-200nm. Occasionally, there will be a large number of filamentous forms with a length of Up to 1000nm. There are four subtypes of avian metapneumovirus, A, B, C, and D. In 2012, we first discovered avian metapneumovirus C infect...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/155A61P31/14C12R1/93
Inventor 韦莉刘爵阎旭朱珊珊全荣王菁侯磊李子璇
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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