A new indicator cell line established to monitor bovine foamy virus infection |
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Authors: | Hong-yan Guo Zhi-bin Liang Yue Li Juan Tan Qi-min Chen Wen-tao Qiao |
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Affiliation: | Key Laboratory of Molecular Microbiology and Biotechnology, Ministry of Education, and Key Laboratory of Microbial Functional Genomics of Tianjin, College of Life Sciences, Nankai University, Tianjin 300071, China |
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Abstract: | In order to improve the accuracy for quantitating the bovine foamy virus (BFV) in vitro, we developed a baby hamster kidney cell (BHK)-21-derived indicator cell line containing a plasmid that encodes the firefly luciferase driven by the BFV long terminal repeat promoter (LTR, from −7 to 1012). The BFV titer could be determined by detecting the luciferase expression since the viral trans-activator BTas protein activates the promoter activity of the LTR. One clone, designated BFVL, was selected from ten neomycin-resistant clones. BFVL showed a specific and inducible dose- and time-dependent luciferase activity in response to BFV infection. Although the changes in luciferase activity of BFVL peaked at 84 h post infection, it was possible to differentiate infected and uninfected cells at 48 h post infection. A linear relationship was established between the multiplicity of infection (MOI) of BFV and the activated ratio of luciferase expression in BFVL. Moreover, the sensitivity of the BFVL-based assay for detecting infectious BFV was 10,000 times higher than the conventional CPE-based assay at 48 h post infection. These findings suggest that the BFVL-based assay is rapid, easy, sensitive, quantitative and specific for detection of BFV infection. |
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Keywords: | Bovine foamy virus Firefly luciferase Indicator cell line |
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