Product-enhanced reverse transcriptase assay for replication-competent retrovirus and lentivirus detection

Lakshmi Sastry, Yi Xu, Lisa Duffy, Sue Koop, Aparna Jasti, Holger Roehl, Doug Jolly, Kenneth Cornetta

Research output: Contribution to journalArticle

27 Scopus citations

Abstract

The product-enhanced reverse transcriptase (PERT) assay has been used to detect reverse transcriptase (RT) activity associated with retroviruses. Although the PERT assay has been proposed as a method for detection of replication-competent retrovirus (RCR) and lentivirus (RCL), it has not been rigorously compared with existing methods for RCR and RCL detection. We have assessed the PERT assay for detection of RCL and RCR that may contaminate lentiviral and retroviral vectors and compared it with published methods for RCL (p24gag ELISA/gag PCR) and RCR (S+/L-) detection. Our results suggest that the PERT assay is as sensitive as p24 gag ELISA and gag PCR for detection of replication-competent HIV-1 in an RCL detection assay. Comparison of detection of replication-competent retroviruses, GALV and RD114, by extended S+/L- and PERT assays indicates that both assays can detect 1 IU of each virus. Our findings suggest that the PERT assay can be used for RCL and RCR testing of a variety of retroviral vectors regardless of the structure, sequence, and envelope of the vectors.

Original languageEnglish (US)
Pages (from-to)1227-1236
Number of pages10
JournalHuman gene therapy
Volume16
Issue number10
DOIs
StatePublished - Oct 1 2005

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

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