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INTRODUCTION: Barr body is of the X chromosomes formed from random inactivation and condensation of one of the two female chromosomes in virtually all the somatic cells of female mammalsAIM: The study aimed to compare the specificity, sensitivity and reliability of May Grunwald-Giemsa (MGG), Carbol fuchsin (CF) and Van Geison (VG) stain in identifying barr bodies in buccal mucosal smears.MATERIALS AND METHODS: A prospective single blinded cohort study was conducted on buccal mucosal smears obtained from 30 subjects. The study subjects was divided into group I and group II consisting of 15 females and 15 males in each group respectively following obtainment of informed consent. All the buccal mucosal smears were processed accordingly as described by Ayyad S B et al (2012), Meena N et al (2016) and Wadsworth A et al (2012) for May Grunwald-Giemsa stain, Carbol fuchsin stain and Van Geison stain. The barr bodies identification was determined by the presence of a darkly stained condensed area on the nucleoplasm as described by Balderman S et al.RESULTS: The results thus obtained showed that CF had a better BBI compared to MGG and VG and the statistical analysis showed 100% specificity, sensitivity and reliability in all the three stains in identifying barr bodies. Barr body was found to be negative in the male samples and positive for female samples with Carbol fuschin yielding 9.8% of Bar Body Index (BBI), May Grunwald-Giemsa yielding 5.4% and Van Geison yielding 2.8%.CONCLUSION: On comparison, the efficacy and accuracy for detection of Barr bodies were more with Carbol fuschin following May Grunwald-Giemsa and Van Geison.