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Radiation Measurements with a DNA Double-Strand-Break Dosimeter

M Obeidat

M Obeidat, K Cline , S Stathakis , N Papanikolaou , K Rasmussen , A Gutierrez , CS Ha , SE Lee , EY Shim* , N Kirby* , University of Texas HSC SA, San Antonio, TX, *Co-Principal Investigators


MO-AB-BRA-4 (Monday, August 1, 2016) 7:30 AM - 9:30 AM Room: Ballroom A

Purpose:Many types of dosimeters are used to measure radiation, but none of them directly measures the biological effect of this dose. The purpose here is to create a dosimeter that can measure the probability of double-strand breaks (DSB) for DNA, which is directly related to the biological effect of radiation.

Methods:The dosimeter has DNA strands, which are labeled on one end with biotin and on the other with fluorescein. The biotin attaches these strands to magnetic beads. We suspended the DNA dosimeter in phosphate-buffered saline (PBS) as it matches the internal environment of the body. We placed small volumes (50μL) of the DNA dosimeter into tubes and irradiated these samples in a water-equivalent plastic phantom with several doses (three samples per dose). After irradiating the samples, a magnet was placed against the tubes. The fluorescein attached to broken DNA strands was extracted (called the supernatant) and placed into a different tube. The fluorescein on the unbroken strands remained attached to the beads in the tube and was re-suspended with 50μL of PBS. A fluorescence reader was used to measure the fluorescence for both the re-suspended beads and supernatant. To prove that we are measuring DSB, we tested dosimeter response with two different lengths of attached DNA strands (1 and 4 kilo-base pair).

Results:The probability of DSB at the dose levels of 5, 10, 25, and 50 Gy were 0.05, 0.08, 0.12, and 0.19, respectively, while the coefficients of variation were 0.14, 0.07, 0.02, and 0.01, respectively. The 4 kilo-base-pair dosimeter produced 5.3 times the response of the 1 kilo-base-pair dosimeter.

Conclusion:The DNA dosimeter yields a measurable response to dose that scales with the DNA strand length. The goal now is to refine the dosimeter fabrication to reproducibly create a low coefficient of variation for the lower doses.

Funding Support, Disclosures, and Conflict of Interest: This work was supported in part by Yarmouk University (Irbid, Jordan) and CPRIT (RP140105)

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