Method for determination and quantification of radiation or genotoxin exposure
The present invention discloses methods for detecting exposure of a living subject to genotoxic agents, testing sensitivity to a genotoxic agent, and determining DNA damage caused by exposure to an agent, comprising detecting the presence of FANCD2-containing foci from a sample collected from said subject. The presence of concentrated foci is indicative of DNA damage, and the degree of foci formation is correlated with degree of exposure. Diagnostic reagents contain a ligand that binds to human FANCD2 associated with a detectable label. Kits for detecting DNA damage in a biological sample contain such diagnostic reagents and signal detection components. The invention further discloses methods for identifying agents which modulate the ability of FANCD2-containing foci to form. Among other things, such agents are potentially useful chemosensitizing agents or may confer protection against damage caused by genotoxic agents.
1. A method of detecting exposure to a genotoxic agent, comprising:
(a) identifying a subject who is at risk of being exposed to said genotoxic agent;
(b) collecting a sample from said subject;
(c) detecting the number of FANCD2-containing foci in said sample;
(d) detecting the number of FANCD2-containing foci in a control sample; and
(e) comparing the number of FANCD2-containing foci detected in (c) with the number of FANCD2-containing foci detected in (d),
wherein if the number of FANCD2-containing foci detected in (c) exceeds the number of FANCD2-containing foci detected in (d), said exposure to said genotoxic agent is detected.
2. The method of claim 1 , further comprising contacting said sample with a ligand which binds to human FANCD2 of SEQ ID NO: 1, wherein said ligand is associated with a label which provides a detectable signal.
3. The method of claim 2 , wherein said ligand comprises a protein that binds to human FANCD2 of SEQ ID NO: 1 and is fused in frame with a fluorophore.
4. The method of claim 1 , further comprising before step (d), a step of exposing said control sample to said genotoxic agent.