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J Virol Methods. 2010 Dec;170(1-2):9-15. doi: 10.1016/j.jviromet.2010.08.004. Epub 2010 Aug 11.

A pseudo-plaque method for infectious particle assay and clonal isolation of adeno-associated virus.

Author information

1
Institute of Molecular Biophysics, Florida State University, Tallahassee, FL 32306-4380, USA.

Abstract

A colorimetric method has been developed for the detection of adeno-associated virus (AAV) infectious centers in cell culture monolayers. Due to its non-cytopathic nature, AAV has not been amenable to the traditional plaque assay, involving an agar overlay and cellular stains. As a result, an alternate method was required. The pseudo-plaque assay is based on enzyme-catalyzed color development after a fixed cell monolayer is probed with anti-AAV monoclonal antibodies. In spite of chemical fixation, expected to damage the viral genomes and particles, infectious particles can be recovered and amplified for the propagation of viral clones.

PMID:
20708035
PMCID:
PMC2981658
DOI:
10.1016/j.jviromet.2010.08.004
[Indexed for MEDLINE]
Free PMC Article

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