An integrated cell culture and quantitative polymerase chain reaction technique for determining titers of functional and infectious adenoviruses.
We describe a simple and rapid assay for determining the titer of functional and infectious adenoviruses that yields reliable results within 36 h. The method consists of an initial incubation of serial diluted adenoviruses on HEK293 cells and a subsequent detection of adenovirus genomic DNA with quantitative polymerase chain reaction. With an adenovirus of known titer as a reference, the exact titers of four unknown adenoviruses could be easily and accurately determined in one 96-well plate. This method can be potentially modified for quality control of other viruses.
Li
Feng
F
Center for Vaccines and Biotherapeutics, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou International Business Incubator A3, Guangzhou Science City, Guangzhou 510663, China. li_feng@gibh.ac.cn
Feng
Liqiang
L
Liu
Yichu
Y
Zhen
Xuehua
X
Chen
Ling
L
D000256:Adenoviridae / Q000737:chemistry / Q000235:genetics* / Q000254:growth & development; D052160:Benzothiazoles; D002460:Cell Line; D003959:Diamines; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D009930:Organic Chemicals / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000379:methods*; D011804:Quinolines; D017186:Titrimetry
D052160:Benzothiazoles; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I