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.
Chang
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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