期刊
JOURNAL OF VETERINARY MEDICAL SCIENCE
卷 70, 期 8, 页码 785-790出版社
JAPAN SOC VET SCI
DOI: 10.1292/jvms.70.785
关键词
detection; endogenous retrovirus; RD114 virus
资金
- Ministry of Education, Culture, Sports, Science and Technology
- Ministry of Agriculture, Forestry and Fisheries
Cats have an infectious endogenous retrovials, named RD114 virus, and there is a possibility that RD114 virus has contaminated live attenuated vaccines, for which feline cells are used as a substrate. To monitor infectious RD114 virus in vaccines for cats, we developed a LacZ marker rescue assay to detect infectious RD 114 virus. Among four human cell lines examined, TE671 cells (human rhabdomyosarcoma) were most susceptible to RD 114 virus and supported RD 114 replication efficiently. Infection was enhanced approximately 5 times by the addition of polybrene at concentrations of 2 to 8 mu g/m/ in the medium during viral adsorption. A 4-hr viral adsorption period was sufficient to obtain the maximum titer. By inoculating samples into TE671 cells transduced with the lacZ marker gene, the limiting diluted sample (i.e., less than 10 infectious units) was detected at 12 clays post-inoculation by the LacZ marker rescue assay. Based on the results obtained in this study, we propose a standard protocol of the LacZ marker rescue assay to detect infectious RD114 virus.
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