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 S. Chapman

doi:10.1016/j.jviromet.2010.08.004

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 S. Chapman

Research output: Contribution to journal › Article › peer-review

5 Scopus citations

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 language	English (US)
Pages (from-to)	9-15
Number of pages	7
Journal	Journal of Virological Methods
Volume	170
Issue number	1-2
DOIs	https://doi.org/10.1016/j.jviromet.2010.08.004
State	Published - Dec 2010
Externally published	Yes

Keywords

AAV
Adeno-associated virus
Assay
Plaque

ASJC Scopus subject areas

Virology

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10.1016/j.jviromet.2010.08.004

Cite this

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

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.",

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AU - Lerch, Thomas F.

AU - Hare, Joan T.

AU - Chapman, Michael S.

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AB - 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.

KW - AAV

KW - Adeno-associated virus

KW - Assay

KW - Plaque

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

Abstract

Keywords

ASJC Scopus subject areas

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