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Purification method for single virus in avian leukemia virus mixed infection

An avian leukemia virus and mixed infection technology, applied in the field of virus purification, can solve problems such as reducing the success rate of vaccine immunization and affecting production performance

Inactive Publication Date: 2017-08-18
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

ALV can cause immunosuppression of the body, reduce the success rate of vaccine immunization, and seriously affect production performance

Method used

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  • Purification method for single virus in avian leukemia virus mixed infection
  • Purification method for single virus in avian leukemia virus mixed infection
  • Purification method for single virus in avian leukemia virus mixed infection

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0018] 1.1 strain

[0019] The avian leukosis virus mixed with subtypes A, B, and J comes from the Chinese isolate GX15MM6-2-Mix strain (preserved by the applicant's laboratory).

[0020] 1.2 Method

[0021] 1.2.1 Primer design

[0022] The primers were designed according to the literature, and the ALV typing primers are shown in Table 1, and were synthesized by Shanghai Huada Bioengineering Co., Ltd.

[0023] Table 1 Primer Sequence

[0024]

[0025]

[0026] 1.2.2 Recovery of strain GX15MM6-2Mix

[0027] Inoculate DF-1 cells with virus strain GX15MM6-2Mix, continue to incubate at 37°C for 1h, discard the supernatant after 1h, and replace with DMEM medium containing 1% fetal bovine serum, at 37°C, 5% CO 2 Conditions were cultivated for 9 days; then the cell culture was collected and stored at -80°C for later use.

[0028] 1.2.3 Determination of TCID50 of strain GX15MM6-2Mix

[0029] Propagate the recovered mixed infection strain GX15MM6-2Mix in DF-1 for 3 generati...

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Abstract

The invention discloses a purification method for a single virus in avian leukemia virus mixed infection. End point dilution is carried out by measuring TCID50 of mixed inflection virus, DF-1 cells are inoculated by adopting a diluted virus solution, supernatant and a cell culture are collected, and a parting authentication primer and an IFA are used for carrying out authentication, so that a single subtype avian leukemia virus is obtained. The method is simple for experiment condition requirement, and has extremely high practical application values. Experimental results prove that different subtype mixed infection ALVs can be purified successfully, and technical support can be provided for research of avian leukemia virus molecular biology characteristics and pathogenicity thereof.

Description

technical field [0001] The invention belongs to the technical field of virus purification, and in particular relates to a method for purifying a single virus in avian leukosis virus mixed infection. Background technique [0002] Avian leukemia virus (ALV) is a retroviridae, leukemia / sarcoma virus group. In 1868, Roloff reported a case of "lymphosarcoma", which was the first report on avian leukaemia. Subsequently, ALV has been reported in various places, and its host range has gradually expanded, spreading from broiler chickens to laying hens and local breed chickens, and its clinical tumor manifestations have become more and more obvious, causing greater economic losses to the poultry industry. [0003] ALV is divided into 7 subgroups: A, B, C, D, E, J, and K. Among them, A, B, and J subgroups are easy to separate clinically, and K subgroup is a new subgroup discovered recently. ALV can cause immunosuppression of the body, reduce the success rate of vaccine immunization, ...

Claims

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Application Information

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IPC IPC(8): C12N7/02C12Q1/70C12Q1/68C12R1/93
CPCC12N7/00C12N2740/11051C12Q1/702
Inventor 王培坤林璐璐杨永立李海娟磨美兰韦天超黄腾韦平
Owner GUANGXI UNIV
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