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A method for isolating avian metapneumovirus

A technology of avian metapneumovirus and isolation method, which is applied in the field of virus isolation and inoculation, can solve problems such as difficulty in obtaining virus isolates, failure to isolate viruses, and inappropriate culture methods, so as to improve in vitro culture conditions, reduce passage times, and improve isolation. rate effect

Active Publication Date: 2018-02-27
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In actual work, it is difficult to obtain the expected virus isolates or fail to isolate viruses due to mixed infection of samples with multiple pathogens, sample storage and transportation conditions, and inappropriate culture methods.

Method used

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  • A method for isolating avian metapneumovirus
  • A method for isolating avian metapneumovirus
  • A method for isolating avian metapneumovirus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Adopt SPF chicken embryo trachea of ​​the present invention, lung primary mixed cell inoculation method and traditional SPF chicken embryo yolk sac inoculation, chicken embryo fibroblast, Vero cell and tracheal ring tissue inoculation method, carry out avian metapneumovirus isolation rate comparison, the same After the samples were processed, the method of the present invention and the traditional method were used to isolate viruses.

[0016] The samples were 51 samples of fresh avian metapneumovirus-infected chickens collected from a chicken farm in Weihai, Shandong in 2013 (positive by RT-PCR). Inoculation according to the present invention, the specific operation method is as follows: the sample processing method is the same as 1 "sample processing" in the "summary of the invention". Cell preparation is the same as 2, 3, 4, 5 in "Contents of the Invention". The sample treatments in the following examples and comparative examples are the same and will not be repeated...

Embodiment 2

[0041] Adopt the method of the present invention and above-mentioned 4 kinds of traditional methods, carry out the comparison of separation rate of avian metapneumovirus, and adopt two kinds of methods to carry out virus separation respectively after the same sample is processed.

[0042] The sample is 79 parts of fresh poultry metapneumovirus infected chicken samples collected from Shandong, Hebei, Liaoning, Jiangsu four provinces in 2014 (detected as positive through RT-PCR), inoculated according to the present invention, specific operation method is as follows: sample processing method Same as 1 "Sample Processing" in "Content of the Invention". Cell preparation is the same as 2, 3, 4, 5 in "Contents of the Invention". The sample treatments in the following examples and comparative examples are the same and will not be repeated here. Then take 1ml of the supernatant of avian metapneumovirus disease samples frozen at -70°C and have been processed, filter and sterilize them ...

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Abstract

The invention provides a method for isolating the avian metapneumovirus. The invention adopts the mixed cells of the trachea and lung tissue of hatched SPF chicken embryos aged 17 to 19 days to culture the avian metapneumovirus. On the basis of the traditional method of separating and culturing one clinically ill sample and one sensitive cell at a time, the present invention successfully improves the in vitro culture conditions of AMPV through the same culture system, the separation and passage method of two mixed cells, and significantly improves the separation of AMPV rate, reducing the number of passages.

Description

technical field [0001] The invention belongs to the technical field of virus isolation and inoculation, and in particular relates to a method for isolating avian metapneumovirus. technical background [0002] Avian metapneumovirus (AMPV), also known as Turkey Rhinotracheitis Virus (TRTV), is a member of Paramyxoviridae, Pneumoviridae, and Pneumoviridae. AMPV infection in poultry can cause acute upper respiratory tract infection, which can cause severe respiratory symptoms in broiler chickens and head swelling in adult chickens. It is the main viral pathogen that induces Swollen Head Syndrome (SHS). The disease was first discovered in the Republic of South Africa in 1978 and has since been reported worldwide. In 1998, my country isolated the virus from chickens with swollen head syndrome in a broiler farm in Heilongjiang. The whole genome of AMPV includes 8 genes, namely N, P, M, F, M2, SH, G, L. According to the difference of viral G protein and serological analysis, AMPV...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00C12R1/93
Inventor 邹敏吴发兴刘新文宫晓范根成孙健
Owner YEBIO BIOENG OF QINGDAO
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