Establishment of an avian leukosis virus subgroup A-resistant cell line

Rapid diagnostic methods for classifying avian leukosis subgroups in the field were needed for routine, large-scale screening. As a first step in method development, we inserted the avian leukosis virus subgroup A （ALV-A） env gene into plasmid pcDNA3.1/Zeo （＋） and used this construct to transfect DF...

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Bibliographic Details
Published in:Journal of Integrative Agriculture 2017-04, Vol.16 (4), p.930-936
Main Authors: FENG, Min, DAI, Man-man, LIAO, Ming, CAO, Wei-sheng
Format: Article
Language:English
Subjects:
avian leukosis virus subgroup A (ALV-A)
DF-1/A cell line
diagnostic tool
ELISA
env
亚型
地方分离株
快速诊断方法
病毒性
禽白血病病毒
细胞系
细胞转染
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Summary:Rapid diagnostic methods for classifying avian leukosis subgroups in the field were needed for routine, large-scale screening. As a first step in method development, we inserted the avian leukosis virus subgroup A （ALV-A） env gene into plasmid pcDNA3.1/Zeo （＋） and used this construct to transfect DF-1 cells. Zeocin-resistant cells were obtained after 2 weeks of zeocin selection. Then, the cells were analyzed using PCR, immunofluorescence, and Western blot for expression of the envA-encoded envelope protein after 30 serial passages. The DF-1/A cell line was completely resistant to 104 TCIDso/0.1 mL （50% tissue culture infective dose）ALV-A and was partially resistant to 10~ TCIDs0/0.1 mL ALV-A viral particles. By comparing the DF-1/A and DF-1 cell lines, an ALV-A isolate was identified using a gag-specific ELISAfor capsid protein p27. Thus, we established a DF-1/A cell line that was resistant to ALV-A infection. This cell line will be useful as a diagnostic tool.
ISSN:2095-3119
2352-3425
DOI:10.1016/S2095-3119(16)61453-3