title: A new indicator cell line established to monitor bovine foamy virus infection.
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.
authors: 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, China.
authors: Hong-Yan
authors: HY
authors: Guo
authors: Zhi-Bin
authors: ZB
authors: Liang
authors: Yue
authors: Y
authors: Li
authors: Juan
authors: J
authors: Tan
authors: Qi-Min
authors: QM
authors: Chen
authors: Wen-Tao
authors: WT
authors: Qiao
mesh_terms: D000818:Animals; D019677:Artificial Gene Fusion; D002417:Cattle; D002418:Cattle Diseases / Q000821:virology*; D002460:Cell Line; D006224:Cricetinae; D005453:Fluorescence; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D012192:Retroviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D012680:Sensitivity and Specificity; D016092:Spumavirus / Q000302:isolation & purification*; D019562:Viral Load / Q000379:methods*
chemical_list: D008156:Luciferases