A quantitative assay for measuring of bovine immunodeficiency virus using a luciferase-based indicator cell line.
In order to quantitate the bovine immunodeficiency virus (BIV) infection in vitro, a BIV indicator cell line (BIVL) was established by transfecting baby hamster kidney cells with reporter plasmids containing the firefly luciferase gene driven by a BIV long terminal repeat promoter. The BIV activates promoter activity of the LTR to express luciferase upon infection. BIV infection could therefore by quantified by detection of luciferase activity. Compared to standard assays used to detect BIV infection, the BIVL-based assay is 10 times more sensitive than the the CPE-based assay, and has similar sensitivity with the viral capsid protein Western blot assay. BIV indicator cell line could detect BIV infection specifically. Luciferase activity of BIV infected BIVL cells showed a time dependent manner, and 60 h post infection is the optimal time to detect BIV infection. Luciferase activity of BIVL cells correlates with the BIV capsid protein expression. Moreover, a linear relationship was found between MOI and the activated intensity of luciferase expression. In brief, the BIV indicator cell line is an easy, robust and quantitive method for monitoring BIV infection.
Yao
Xue
X
Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Nankai University, Tianjin 300071, China.
Guo
Hong-yan
HY
Liu
Chang
C
Xu
Xuan
X
Du
Jian-sen
JS
Liang
Hao-yue
HY
Geng
Yun-qi
YQ
Qiao
Wen-tao
WT
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D017930:Genes, Reporter; D016087:Immunodeficiency Virus, Bovine / Q000302:isolation & purification*; D049409:Luciferases, Firefly / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D012680:Sensitivity and Specificity; D019562:Viral Load / Q000379:methods*
D049409:Luciferases, Firefly