Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 114, Issue 1, Pages 55-64Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2003.08.012
Keywords
real-time polymerase chain reaction; p53 recombinant adenovirus; gene therapy; cancer
Ask authors/readers for more resources
The purpose of this study was to assess the usefulness of real-time PCR as a quantitative, highly reproducible, and sensitive method, for detecting and quantifying p53 recombinant adenovirus in biological samples from cancer patients receiving injections of Ad5CMV-p53. The dynamic range of this real-time PCR-based assay was wide (at least five orders of magnitude). Our assay used an internal positive control in the same PCR tube that is capable of detecting residual PCR inhibitors. Serial spiked samples in plasma with known quantities of Ad5CMV-p53 were evaluated. The minimum detection limit was 2 pfu per PCR (similar to50 pfu per ml of plasma) and the quantification values were reproducible. A total of 2069 controls tested with 1780 plasma samples from 286 patients enrolled in gene therapy trials using Ad5CMV-p53 were investigated. Using calibrators to adjust the quantitation value, the results confirmed the good performance of the assay. In conclusion, the high sensitivity, simplicity and reproducibility of the real-time Ad5CMV-p53 assay, allowing screening of large numbers of samples, combined with its wide dynamic range, make this method particularly suitable for monitoring gene therapy trials. (C) 2003 Elsevier B.V. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available