Establishment of a LacZ Marker Rescue Assay to Detect Infectious RD114 Virus

• DOI: 10.1292/jvms.70.785
• Publication Date: 2008-09-05T06:03:45Z
• Authors: Shoichi SAKAGUCHI; Masaya OKADA; Takayuki SHOJIMA; Kenji BABA; Takayuki MIYAZAWA
• Keywords: Endogenous retrovirus

Cats have an infectious endogenous retrovirus, named RD114 virus, and there is a possibility that virus has contaminated live attenuated vaccines, for which feline cells are used as substrate. To monitor in vaccines cats, we developed LacZ marker rescue assay to detect virus. Among four human cell lines examined, TE671 (human rhabdomyosarcoma) were most susceptible supported replication efficiently. Infection was enhanced approximately 5 times by the addition of polybrene at concentrations 2 8 μg/ml medium during viral adsorption. A 4-hr adsorption period sufficient obtain maximum titer. By inoculating samples into transduced with lacZ gene, limiting diluted sample (i.e., less than 10 units) detected 12 days post-inoculation assay. Based on results obtained this study, propose standard protocol