Gamma-X satellite DNA is a 220-bp tandemly arranged repetitive DNA with specificity for the centromeric region of the human X chromosome. The conservation of this human X centromeric satellite DNA sequence in primate species was evaluated by comparative fluorescence in-situ hybridization to metaphase chromosome preparations of the great apes and three Old World monkeys. Homologous gamma-X DNA were detected at centromeric locations in all six primate species. For the great apes, gamma-X was exclusively localized to the centromeric regions of the X chromosomes. Among the Old World monkeys studied, only the golden monkey exhibited localization to the X chromosome. In the black-and-white colobus and the pig-tailed macaque, human gamma-X sequences were localized to the pericentromeric regions of autosomes 1 and 4, respectively.