Capability for identification of gamma-irradiated bovine liver by new high sensitivity comet assay.
DNA in food will sustain damage by gamma radiation. The detection capability of the high sensitivity comet assay was studied using fluorescence-microscopy. Beef liver was irradiated at a range of 1 Gy to 8 kGy. Single cells were obtained from the irradiated liver, then analyzed by agaros-gel electrophoresis. The pH of the buffer for electrophoresis was pH 13, which is generally utilized for sensitive detection of DNA damage. The pattern formed by DNA was visualized by staining with ethidium bromide. The resulting comets were evaluated with a scale we developed, and Influence Scores were calculated based on the Tice method. It is possible to detect irradiation damage to beef liver at 10 Gy. Together with Influence Score, histogram of comet type is used for detection of irradiation. We elucidated those histograms were useful for distinguishing damage caused by irradiation from that of others. DNA damage can be caused not only by irradiation, but also by the other treatments. Therefore, the respective influences of freezing, preservation, irradiating temperature, atmosphere of irradiation, cooking, and homogenizing devices were also examined. This new comet assay will be a useful method of detecting DNA damage to identify irradiated foods.
Miyahara
M
M
National Institute of Health Sciences, Tokyo, Japan. mmiyaha@nihs.go.jp
Saito
A
A
Ito
H
H
Toyoda
M
M
D000818:Animals; D001272:Atmosphere; D002417:Cattle; D003080:Cold Temperature; D020552:Comet Assay / Q000379:methods*; D003296:Cooking; D004249:DNA Damage; D004307:Dose-Response Relationship, Radiation; D005504:Food Analysis; D005511:Food Handling; D005514:Food Irradiation; D005519:Food Preservation; D005720:Gamma Rays; D008099:Liver / Q000528:radiation effects*; D008856:Microscopy, Fluorescence; D012680:Sensitivity and Specificity