Patent ID: 7824874

Claim:
A method of assessing the mutagenic DNA-damaging potential of an agent comprising: providing a blood or bone marrow sample from a mammal exposed to an agent that may induce mutagenic DNA damage, the blood or bone marrow sample comprising erythrocytes and reticulocytes; first contacting the sample with a first reagent that binds GPI-anchor-expressing erythrocytes and reticulocytes, but not GPI-anchor-deficient erythrocytes and reticulocytes, wherein the first reagent comprises a first fluorochrome; second contacting the sample with a second reagent that binds to mammalian platelets, wherein the second reagent comprises a second fluorochrome having a fluorescent emission spectrum that does not substantially overlap with a fluorescent emission spectrum of the first fluorochrome; third contacting the sample with a third fluorochrome-containing reagent that labels reticulocytes but not mature erythrocytes, the third fluorochrome having a fluorescent emission spectrum that does not substantially overlap with the fluorescent emission spectra of the first and second fluorochromes; exciting the first, second, and third fluorochromes with light having a wavelength suitable to induce fluorescence thereof; and detecting the fluorescent emission and light scatter produced by erythrocytes and reticulocytes labeled with the first fluorochrome, while excluding platelets labeled by the second fluorochrome and nucleated cells but not reticulocytes labeled by the third fluorochrome, and counting the number of GPI-anchor deficient erythrocytes and/or reticulocytes relative to the number of total erythrocytes and/or reticulocytes, wherein a statistically significant deviation in the frequency of GPI-anchor-deficient erythrocytes and/or reticulocytes from a baseline frequency of GPI-anchor-deficient erythrocytes and/or reticulocytes, respectively, in unexposed or vehicle control mammals indicates the mutagenic DNA-damaging potential of the agent.