Effects of Shipping and Handling On a DNA-Based Detector
T Schulze1*, H Nusrat2, G Pang3, M Tambasco4, (1) SDSU, San Diego, CA, (2) Sunnybrook Health Sciences Center, Toronto, ON, (3) Odette Cancer Centre, Toronto, ON, CA, (4) San Diego State Univ, San Diego, CA
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MO-IePD-TRACK 6-6 (Monday, 7/26/2021) 5:30 PM - 6:00 PM [Eastern Time (GMT-4)]
Purpose: Our lab has developed a DNA-based detector that uses plasmid DNA as a biosensor of DNA damage from ionizing radiation. We aimed to quantify the effects of shipping and handling on the stability of DNA detector readings to assess radiobiological damage.
Methods: DNA detectors were constructed by pipetting plasmid DNA, pBR322 (Thermo Fisher Scientific, Inc.), into nine 6.28 mm³ cylindrical wells drilled into biocompatible near tissue equivalent material (polycarbonate). Each well contained 3 μl of DNA solution with a concentration of 12 ng/μl for a total of 27 μl per detector. Each detector was sealed using aluminum foil seals. One control and two detectors to be irradiated to 20 and 30 Gy, respectively, using the XStrahl 300 orthovoltage unit, were shipped on ice in an insulated box via FedEx airmail from San Diego State University (SDSU), San Diego, USA, to Odette Cancer Centre Sunnybrook, Toronto, Canada following biomaterials shipment guidelines. A stationary control was kept at -20 ℃ at SDSU. The detectors were shipped back to SDSU via FedEx airmail. Agarose gel electrophoresis was used to separate the plasmid DNA open circular, linear, and supercoiled isoform bands corresponding to single strand breaks (SSBs), double strand breaks (DSBs), and no strand breaks, respectively. The gel band intensities were imaged to quantify the relative SSBs and DSBs.
Results: Total shipment distance and time was 8300 km in ~3.6 days. The shipped controls resulted in mean increases of 18.3±3.0% and 1.64±0.30% in the SSBs and DSBs, respectively. Adjusted for these changes, the 20 and 30 Gy irradiated samples showed DNA DSB damage of 4.73±1.10% and 8.26±0.67%, respectively.
Conclusion: Increases in SSBs and DSBs occurred with shipping and handling of the DNA. However, relative damage between the shipped and stationary control can be used to quantify DNA damage in the irradiated detectors.
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