The present study describes a rare case of freemartinism of the 60XX/60XY chimera type in an adult 7-year-old horned, mixed Lori breed phenotypically doe. Signs of masculinisation were evident in both the behavioral patterns and body conformation, but there was no structure similar to penis and hypospadias was observed in this goat. External genital organs were similar to that of a female one. In this study, a single multiplex PCR assay was used to determine the freemartinism status (XX-XY blood chimerism) based on semi-quantitatively co amplify a sex-based polymorphism in the amelogenin locus (AMX and AMY). In addition, serum concentrations of anti-Mullerian hormone (AMH), testosterone and progesterone hormones of this horned intersex goat were evaluated. Serum AMH levels in the present case (0.2 ng/ml) were similar to male (0.2 ng/ml) and lower than female (0.6 ng/ml) goat. The serum concentrations of testosterone (0.2 ng/ml) were remarkably high in comparison to normal control male and female ones (0.02 ng/ml). Progesterone level in the intersex goat (0.4 ng/ml) was close to control buck (0.7 ng/ml) and remarkably lower than control doe (10 ng/ml). The results of the present study showed that the use of hormonal pattern, especially AMH level, and single multiplex PCR technique provide an easy and alternative approaches for indication of XX/XY chimerism or mosaicism in intersex freemartin goats.