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

Daniel A J Mitchell, Thomas F. Lerch, Joan T. Hare, Michael Chapman

Research output: Contribution to journalArticle

4 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)9-15
Number of pages7
JournalJournal of Virological Methods
Volume170
Issue number1-2
DOIs
StatePublished - Dec 2010

Fingerprint

Dependovirus
Viral Genome
Virion
Agar
Coloring Agents
Clone Cells
Color
Cell Culture Techniques
Monoclonal Antibodies
Enzymes

Keywords

  • AAV
  • Adeno-associated virus
  • Assay
  • Plaque

ASJC Scopus subject areas

  • Virology

Cite this

A pseudo-plaque method for infectious particle assay and clonal isolation of adeno-associated virus. / Mitchell, Daniel A J; Lerch, Thomas F.; Hare, Joan T.; Chapman, Michael.

In: Journal of Virological Methods, Vol. 170, No. 1-2, 12.2010, p. 9-15.

Research output: Contribution to journalArticle

Mitchell, Daniel A J ; Lerch, Thomas F. ; Hare, Joan T. ; Chapman, Michael. / A pseudo-plaque method for infectious particle assay and clonal isolation of adeno-associated virus. In: Journal of Virological Methods. 2010 ; Vol. 170, No. 1-2. pp. 9-15.
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