Assessing a new gene expression analysis technique for radiation biodosimetry applications

The response to any radiation accident or incident involving actual or potential ionising radiation exposure requires accurate and rapid assessment of the doses received by individuals. The techniques available today for biodosimetry purposes are not fully adapted to rapid high-throughput measurements of exposures in large numbers of individuals. A recently emerging technique is based on gene expression analysis, as there are a number of genes which are radiation responsive in a dose-dependent manner. The present work aimed to assess a new technique which allows the detection of the level of expression of up to 800 genes without need of enzymatic reactions. In order to do so, human peripheral blood was exposed ex vivo to a range of x-ray doses from 5 mGy to 4 Gy of x-rays and the transcriptional expression of five radiation-responsive genes PHPT1, PUMA, CCNG1, DDB2 and MDM2 was studied by both the nCounter Digital Analyzer and Multiplex Quantitative Real-Time Polymerase Chain Reaction (MQRT-PCR) as the benchmark technology. Results from both techniques showed good correlation for all genes with R² values ranging between 0.8160 and 0.9754. The reproducibility of the nCounter Digital Analyzer was also assessed in independent biological replicates and proved to be good. Although the slopes of the correlation of results obtained by the techniques suggest that MQRT-PCR is more sensitive than the nCounter Digital Analyzer, the nCounter Digital Analyzer provides sensitive and reliable data on modifications in gene expression in human blood exposed to radiation without enzymatic amplification of RNA prior to analysis.