Practical assay issues with the PERT/PBRT assay: a highly sensitive reverse transcriptase assay.
Product safety testing for retroviruses can be achieved by a panel of screening assays, including electron microscopy, viral gene specific PCRs, virus propagation, and detection of reverse transciptase activity. The application of PCR-based reverse transcriptase assays (PERT) that are approximately a million-fold more sensitive than conventional nucleotide incorporation assays in the testing of biologicals is described. Use of PERT assays can be applied to three areas: (i) screening for adventitious retrovirus contamination; (ii) detecting and quantifying endogenous viral particle load and (iii) monitoring levels of infectious retrovirus generation in cell lines that contain endogenous retroviruses.
Biologics Testing, BioReliance, Invitrogen Bioservices, Rockville, MD 20850, USA. achang@bioreliance.com
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D000818:Animals; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000592:standards*; D002460:Cell Line; D006224:Cricetinae; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics; D012194:RNA-Directed DNA Polymerase / Q000032:analysis*; D012190:Retroviridae / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013698:Templates, Genetic
D012367:RNA, Viral; D012194:RNA-Directed DNA Polymerase