Identification of adeno-associated virus contamination in cell and virus stocks by PCR.
To further understand the biology of adeno-associated virus (AAV) and identify the presence of AAV in laboratory samples, we have developed a sensitive PCR-based assay using degenerate primers based on the sequence of seven diverse AAV isolates. Using these primers, we can detect free virus in viral stocks, cleared cell lysate, as well as in latently infected cells. The method can detect as little as 10 viral copies/microL of sample and can be adapted for high-throughput screening technology. With this method, we have also detected a new AAV isolate from a stock of bovine adenovirus.
Katano
Hisako
H
National Institutes of Health, Bethesda, MD, USA.
Afione
Sandra
S
Schmidt
Michael
M
Chiorini
John A
JA
D001483:Base Sequence; D045744:Cell Line, Tumor / Q000821:virology*; D017931:DNA Primers; D000229:Dependovirus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers