After allogeneic stem cell transplantation, dual donor and recipient populations may be present. Donor/recipient ratio changes over time may predict clinical outcome: accurate measurement of these changes are needed. Chimerism may be measured by XY-fluorescence in situ hybridization for donor/recipient sex mismatch, or polymerase chain reaction amplification of short tandem repeat loci with donor/recipient sex match. Patients were monitored by each method. Additionally, mononuclear cells from 2 sex-mismatched individuals were mixed and analyzed using both methods. Each gave concordant estimates of patient chimerism and discriminated cell population ratios in mixed blood. We conclude that cytogenetic and molecular methods give accurate donor chimerism estimates.