Chicken Newcastle disease virus and its separation method

A chicken Newcastle disease virus and virus isolation technology, applied in the field of animal virology, can solve the problem that traditional vaccine strains cannot produce effective protection

Inactive Publication Date: 2012-11-07
秦卓明 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] It can be seen that this pathogenic strain and the traditional vaccine strains (La Sota strain and Clone30 strain) all have relatively large differences in genotyping and antigenicity, making the traditional vaccine strains unable to produce effective protection. In order to be able to To better understand the strain and its pathogenicity, it must be further isolated and studied

Method used

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  • Chicken Newcastle disease virus and its separation method
  • Chicken Newcastle disease virus and its separation method
  • Chicken Newcastle disease virus and its separation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The isolation and cultivation of embodiment 1 virus:

[0042] Aseptically collect the trachea, brain, fallopian tube and liver of the diseased chicken, weigh them and add sterilized normal saline in a weight ratio of 1:3, homogenate, ultrasonicate, centrifuge at 3000r / min for 15min, take the supernatant and add green (Strep)mycin double antibody to 2000IU / ml, put in 4°C refrigerator overnight, after passing the sterility test, inoculate five 10-day-old SPF chicken embryos, 0.2ml per embryo. Discard the chicken embryos that died within 24 hours, and then light the eggs once every 6-8 hours, take out the dead chicken embryos in time, refrigerate them at 2-8°C, take out all the chicken embryos until 96 hours, and collect the chicken embryo allantois aseptically After the reserved sample is tested, the virus isolate can be stored at -70°C for future use.

Embodiment 2

[0043] The identification of embodiment 2 virus:

[0044] 1. Hemagglutination test (HA)

[0045] The virus isolate undergoes agglutination reaction to 1% chicken red blood cells, HA is 2 9 , that is, the agglutination titer of the virus is 2 9 .

[0046] 2. Hemagglutination inhibition test (HI)

[0047] The ability of the virus to agglutinate red blood cells can be inhibited by the corresponding specific antibody, namely the hemagglutination inhibition test (HI), which is specific. The HI cross-inhibition test was carried out between virus isolates and specific Newcastle disease, avian influenza H9 and AIV H5 subtypes, Egg Drop Syndrome (EDS) and other viral single-factor positive sera for preliminary differential diagnosis. Specific method: According to the results of the HA test, determine the hemagglutination value of the virus, and prepare a virus solution with 4 hemagglutination units; carry out on a 96-well micro-reaction plate, and use the method of diluting serum w...

Embodiment 3

[0059] Example 3 Comparison of Antigenicity of SDM01 Strain and Other Newcastle Disease Isolates

[0060] 1. HI homology comparison

[0061] For the preparation of single-factor positive serum, according to the national SPF chicken microbial monitoring method, the prepared 20 strains of NDV antigen and positive serum were respectively subjected to cross HI experiment, and the average of 5 detection values ​​was taken. At the same time, set: SPF chicken negative serum, Newcastle disease positive serum control and virus control. The calculation of HI antigen homology is carried out according to the immunological method: r1 = heterologous serum titer 1 / homologous serum titer 1, r2 = heterologous serum titer 2 / homologous serum titer 2, the results are shown in Table 1.

[0062] La Sota had the highest antigenic homology with Clone30 and F48E9, 0.97 and 0.98, respectively, and the antigenic homology with 17 isolates was 0.70-0.97, most of which were between 0.85-0.90. Among the...

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Abstract

The invention relates to the field of animal virology and provides a chicken Newcastle disease virus and its separation method. The virus is code-named SDM01 and the invention provides its separation and preparation method. The inventor carries out biological collection for the strain and its collection number is CCTCC NO:V201109. Through chicken embryo passage, biological characteristics tests such as a hemagglutinin and serum neutralization test, an animal regress test and exogenous virus testing, are carried out. The test results confirm that the virus is the Newcastle disease virus. It isproved through chicken embryo mean death time (MDT), one-day-old chick intracebral pathogenicity index (ICPI) measurement and 6-week-old chick intravenous inoculation pathogenicity index (IVPI) measurement that the Newcastle disease virus is a virulent strain. Positive serum is prepared through separated strain and standard strain to carry out an HI cross neutralization test, a chicken embryo neutralization test, a cell neutralization test, an F and HN gene sequence sequencing and comparison and an immunity protective test. Results confirm that there exist large differences between the Newcastle disease virus SDM01 strain and traditional strains (La Sota strain and Clone30 strain) in both genetic typing and antigenicity.

Description

technical field [0001] The invention relates to the field of animal virology, and specifically provides a new chicken Newcastle disease virus, and provides an isolation method and application of the virus. Background technique [0002] Newcastle disease (NewCastledisease) is a highly contagious infectious disease caused by paramyxovirus. Also known as Asian chicken plague or pseudo chicken plague. Often presents symptoms of acute septicemia Newcastle disease chicken Newcastle disease. The main features are dyspnea, loose stools, nervous disturbances, and mucosal and serosal hemorrhages. The mortality rate is high, and the poultry industry is seriously damaged. There are two types of strong and weak strains. Viruses are divided into three types: low virulence type (i.e. slow onset), moderate virulence type (i.e. medium onset type), and strong virulence type (i.e. rapid onset type). Vaccination is now generally used to prevent such infectious diseases. The most commonly us...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/17A61P31/14C12R1/93
Inventor 秦卓明徐怀英王友令
Owner 秦卓明
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