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DOE OSTI · 1770530

Predicting Ionizing Radiation Exposure (LDRD Final Report)

Dugan, Lawrence C.
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Abstract

Rapid, low-cost, easy-to-use devices are needed for screening large populations potentially exposed to ionizing radiation following accidental or intentional release of radioactive materials. Measurements of radiation responsive messenger RNA (mRNA) biomarkers using fieldable devices could provide such a tool. mRNA expression level changes may be useful for discriminating between exposed individuals and the worried well and estimating the radiation dose received even days after exposure. Loop mediated isothermal amplification, or LAMP, is an established nucleic acid amplification technique that provides key advantages for use in low cost, portable instruments. We proposed to use LAMP to determine if the expression level of one or a small number of radiation responsive biomarkers can be used to discriminate unexposed cells from ionizing radiation (IR)-exposed cells. If successful, this methodology could potentially be transitioned to portable detection technology for use in population screening after large-scale events. Our approach involved designing quantitative, real-time, reverse transcriptase loop-mediated isothermal amplification (RTLAMP) assays for a panel of published IR-responsive mRNA biomarkers and control genes. This assay panel was then tested on mRNA isolated from white blood cells, lymphocyte cell lines and samples from internally dosed individuals. We used standard curves to quantify mRNA copy number along with the comparative Ct method to measure relative mRNA expression changes. We were able to generate quantitative and comparative gene expression changes for many of the samples evaluated. Although promising in its potential to measure gene expression, the technique suffered when attempting to amplify target genes with very low background RNA expression levels, in particular, FDXR in controls samples. Future work should focus on optimizing the RT-LAMP reaction for low RNA expression level targets allowing for the generation of validated assay panels.

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59 BASIC BIOLOGICAL SCIENCES63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.

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BibTeXRIS

Dugan, Lawrence C.. 2019-10-31. Predicting Ionizing Radiation Exposure (LDRD Final Report). https://doi.org/10.2172/1770530

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