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A method for efficiently amplifying subgroup j avian leukosis virus

A technology of avian leukosis virus and subgroups, applied in the field of immunology, can solve the problems of time-consuming, etc., and achieve the effect of improving sensitivity and shortening the separation and detection time

Active Publication Date: 2020-12-29
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although this method is highly reliable, it is time-consuming

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  • A method for efficiently amplifying subgroup j avian leukosis virus
  • A method for efficiently amplifying subgroup j avian leukosis virus

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Experimental program
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Embodiment Construction

[0021] The present invention will be further described below in conjunction with specific examples.

[0022] 1) Cell culture

[0023] Chicken hepatocytes were cultured in DMEM medium containing 10% fetal bovine serum, and DF-1 cells were cultured in DMEM medium containing 5% fetal bovine serum. After the cells covered the monolayer, they were washed once with PBS. Digest with 0.25% trypsin and pass to 6-well or 96-well cell culture plate.

[0024] 2) Efficient amplification of subgroup J avian leukemia virus in chicken hepatocytes

[0025] Chicken hepatocytes and DF-1 cells were inoculated into a 6-well plate for culture. When the confluence was about 70%, the J subgroup avian leukosis virus GY03 strain was inoculated into the two kinds of cells, and the MOI was 0.001. Repeatedly, cultivated with 2ml medium containing 2% fetal bovine serum for 7 days, collected 200 μl supernatant every day and stored it in a -80°C refrigerator for later use, and supplemented 200 μl medium co...

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Abstract

The invention relates to a method for efficiently amplifying the J subgroup avian leukemia virus. The present invention can efficiently amplify the J subgroup avian leukemia virus by using chicken liver cells. The detection time of J subgroup avian leukemia virus can realize early detection of the pathogen and improve the sensitivity of the J subgroup avian leukemia virus detection method. When the MOI of J subgroup avian leukemia virus infection was 0.001, the effect of ELISA in detecting the infection of chicken hepatocytes maintained for 3 days was comparable to that of DF-1 cells maintained for 7 days. The invention establishes a high-efficiency amplification method, which will speed up the purification process of the J subgroup avian leukemia virus, save time and cost, and has good application value in the purification and detection of the J subgroup avian leukemia virus.

Description

technical field [0001] The invention relates to the field of immunology, in particular to a method for efficiently amplifying J subgroup avian leukosis virus. Background technique [0002] Avian leukemia virus (ALV) is a retrovirus that can cause a variety of neoplastic immunosuppressive diseases in poultry. The virus can be divided into 10 subgroups A-J according to the characteristics of its envelope protein, and the A, B, C, D and J subgroups can cause chickens to suffer from different types of tumors of avian leukemia virus, these viruses have vertical and horizontal transmission Ability. Among them, C and D subgroups are very rare. At present, the J subgroup is the most common in China, followed by the A subgroup and the B subgroup. Diseased chickens often show a decrease in egg production rate, emaciation, discarded chicken carcasses, and death of chickens, which lead to huge economic losses in the domestic and even worldwide poultry industry. Because there is no s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00C12N5/071
Inventor 叶建强李拓凡孙舒邵红霞秦爱建谢菁吕璐
Owner YANGZHOU UNIV