A pseudo-plaque method for infectious particle assay and clonal isolation of adeno-associated virus.
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.
Mitchell
Daniel A J
DA
Institute of Molecular Biophysics, Florida State University, Tallahassee, FL 32306-4380, USA.
Lerch
Thomas F
TF
Hare
Joan T
JT
Chapman
Michael S
MS
D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D003124:Colorimetry; D000229:Dependovirus / Q000302:isolation & purification* / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D007124:Immunoenzyme Techniques; D050296:Microbial Viability; D016133:Polymerase Chain Reaction; D019562:Viral Load; D010948:Viral Plaque Assay / Q000379:methods*; D014771:Virion / Q000254:growth & development / Q000302:isolation & purification; D014779:Virus Replication
D000911:Antibodies, Monoclonal