Micronucleus assay with genomic DNA hybridization probe and enzymatic color pigment detection

A method for detecting the presence or absence of micronuclei in anucleated young polychromatic erythrocytes (PCEs) and normochromatic erythrocytes (NCEs) of an organism in a sample, comprising

• (a) isolating PCEs and NCEs of the organism,
• (b) exposing the PCEs and NCEs to a hybridization probe, the hybridization probe comprising digested, labeled whole genomic DNA, the digested genomic DNA being labeled with a first binding member capable of specifically binding with a second binding member,
• whereby, as a result of exposing the PCEs and NCEs to the hybridization probe, the hybridization probe hybridizes with DNA contained in PCEs and NCEs as micronuclei, if present,
• (c) exposing the PCEs and NCEs to a compound comprising the second binding member coupled to an enzyme capable of reacting with a chromogenic substrate
• whereby, as a result of exposing the PCEs and NCEs to the compound, the compound binds to the hybridization probe that is hybridized with the DNA in the PCEs and NCEs as micronuclei, if present,
• (d) exposing the PCEs and NCEs to the chromogenic substrate, whereby the chromogenic substrate is converted into a colored pigment by reaction with the enzyme,
• (e) examining the PCEs and NCEs for the presence of colored pigment, the presence of colored pigment indicating the presence of micronuclei in the PCEs and NCEs, and
• (f) scoring the sample according to the presence or absence of micronuclei in the PCEs and NCEs.