FLUOROMETRIC METHOD FOR RAPID DETECTION OF DNA STRAND BREAKS IN HUMAN WHITE BLOOD-CELLS PRODUCED BY LOW-DOSES OF RADIATION

  • 1 January 1981
    • journal article
    • research article
    • Vol. 41  (5) , 1889-1892
Abstract
DNA strand breaks can be detected with great sensitivity by exposing crude cell lysates to alkaline solutions and monitoring the rate of strand unwinding. As little as 1 strand break per chromosome can be detected. Previous methods for measuring strand unwinding have required physical separation of single- from double-stranded molecules. Conditions are described under which unwinding can be monitored directly using a fluorescent dye, thus greatly simplifying the analysis. Breaks due to irradiation of blood samples by 60Co .gamma.-rays at doses as low as 0.05 to 0.1 gray (5 to 10 rad) were detectable. Rapid rejoining strand breaks during in vitro incubation at 37.degree. C could readily be observed following a dose of 1 gray. Since the procedure is very rapid and cells can be analyzed directly without the requirement for culturing or radiolabeling, the procedure could be useful in cancer chemotherapy if in vivo damage is to be monitored or for testing the in vitro sensitivity of cells to drugs.

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