A new indicator cell line established to monitor bovine foamy virus infection
A new indicator cell line established to monitor bovine foamy virus infection
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建立监测牛泡沫病毒感染的新指示细胞系
DOI：
10.1007/s12250-011-3204-y
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发表时间：
2011-10-01
影响因子：
5.5
通讯作者：
Qiao, Wen-tao
中科院分区：
文献类型：
--
作者：
Guo, Hong-yan;Liang, Zhi-bin;Qiao, Wen-tao
为了提高体外定量检测牛泡沫病毒（BFV）的准确性，我们建立了一种幼仓鼠肾细胞（BHK）-21-源指示细胞系，该细胞系含有编码萤火虫荧光素酶的质粒，该荧光素酶由BFV长末端重复序列启动子（LTR，从-7到1012）驱动。BFV滴度可以通过检测荧光素酶表达来确定，因为病毒反式激活因子BTas蛋白激活LTR的启动子活性。从10个新霉素抗性克隆中选出一个克隆，命名为BFVL。BFVL对BFV感染表现出特异性和诱导性的剂量和时间依赖性荧光素酶活性。虽然BFVL的荧光素酶活性的变化在感染后84 h达到峰值，但在感染后48 h可以区分感染和未感染的细胞。BFV的感染复数（MOI）与BFVL中荧光素酶表达的活化率之间建立了线性关系。此外，在感染后48小时，用于检测感染性BFV的基于BFVL的测定的灵敏度比基于CPE的常规测定高10，000倍。这些结果表明，基于BFVL的检测方法是快速，简便，灵敏，定量和特异性检测BFV感染。
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.