title: Rapid determination of adenoviral vector titers by quantitative real-time PCR.
abstract: Replication defective adenoviruses have been used as vectors in a variety of settings including gene transfer, gene manipulation, and functionality studies. A quantitative real-time PCR-based assay is described for rapid determination of physical titers of recombinant adenovirus vectors. This method is based on amplification of a 77 bp fragment located near the left end of the adenovirus type 5 genome. Evaluation of this method demonstrated that it is simple, sensitive and reproducible, and has a dynamic range of quantitation over 5 logs. This assay is applicable to purified adenovirus as well as vectors prepared by simple cell lysis procedure, requiring only a small amount of starting material. The simplicity and short turn-around time of this assay should facilitate rapid titer determination for a large collection of adenoviral vectors.
authors: Galapagos Genomics BV, Archimedesweg 4, PO Box 2048, 2301 CA Leiden, The Netherlands. libin@galapagos.nl
authors: L
authors: L
authors: Ma
authors: H A
authors: HA
authors: Bluyssen
authors: M
authors: M
authors: De Raeymaeker
authors: V
authors: V
authors: Laurysens
authors: N
authors: N
authors: van der Beek
authors: H
authors: H
authors: Pavliska
authors: A J
authors: AJ
authors: van Zonneveld
authors: P
authors: P
authors: Tomme
authors: H H
authors: HH
authors: van Es
mesh_terms: D000256:Adenoviridae / Q000302:isolation & purification*; D002460:Cell Line; D003199:Computer Systems; D005822:Genetic Vectors / Q000032:analysis*; D016133:Polymerase Chain Reaction / Q000379:methods; D014162:Transfection