A high-throughput hybridization method for titer determination of viruses and gene therapy vectors.
One of the challenges facing researchers working with viruses and gene therapy vectors is the need to rapidly assay for infectious virus. Current methods used to titer many viruses are cumbersome and are not amenable to handling large numbers of samples. Here we describe the development of an assay that can rapidly quantify infectious viruses and gene therapy vectors. The assay relies on biological amplification of viral sequences and hybridization of labeled probes to immobilized nucleic acid from infected cells. The amplification of the viral genome makes this a highly sensitive method. The assay is configured in a high-throughput format that has been used to detect recombinant adeno-associated virus (AAV), wild-type AAV and infectious adenovirus. The assay is quantitative, and can be used to titer virus preparations with or without a known standard.
Atkinson
E M
EM
Development Department, Targeted Genetics Corporation, 1100 Olive Way, Suite 100, Seattle, WA 98101, USA. atkinson@targen.com
Debelak
D J
DJ
Hart
L A
LA
Reynolds
T C
TC
D000260:Adenoviruses, Human / Q000302:isolation & purification*; D002460:Cell Line; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D000229:Dependovirus / Q000302:isolation & purification*; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000302:isolation & purification*; D006367:HeLa Cells; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods*
C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator