Real-time quantitative PCR for the design of lentiviral vector analytical assays.
From the recent and emerging concerns for approving lentiviral vector-mediated gene transfer in human clinical applications, several analytical methods have been applied in preclinical models to address the lentiviral vector load in batches, cells or tissues. This review points out the oldest generation methods (blots, RT activity, standard PCR) as well as a full description of the newest real-time quantitative PCR (qPCR) applications. Combinations of primer and probe sequences, which have worked in the lentiviral amplification context, have been included in the effort to dress an exhaustive list. Also, great variations have been observed from interlaboratory results, we have tempted to compare between them the different analytical methods that have been used to consider (i) the titration of lentiviral vector batches, (ii) the absence of the susceptible emerging replicative lentiviruses or (iii) the lentiviral vector biodistribution in the organism.
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Genethon, CNRS UMR 8115, 1bis rue de l'Internationale, Evry Cedex, France.
Gaillard
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D000818:Animals; D001709:Biotechnology; D002460:Cell Line; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008164:Luminescent Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014161:Transduction, Genetic / Q000379:methods; D019076:Transgenes; D019562:Viral Load
D017931:DNA Primers; D004279:DNA, Viral; D008164:Luminescent Proteins