Aceto-orcein squash technique for barr body detection

Abstract

BACKGROUND: Gender determination is an important aspect of forensic medicine. This can be done by identifying the presence of “Barr body”, which is the inactive ‘X’ chromosome seen in the nucleus at the nuclear membrane of a cell only in females. Sex determination can be done by other methods like polymerase chain reaction and karyotyping, which are expensive and not available in all places. Detection of Barr bodies in buccal smear is easy, cost effective and can be performed routinely with various staining methods.The aims of this study are 1) to identify Barr Bodies for gender determination. 2) To compare the efficacy of aceto-orcein technique and papanicolaou stain for detection of Barr bodies in buccal smears. MATERIAL & METHODS: In this cohort analytic study, buccal smears were prepared from 250 healthy individuals (125 males & 125 females). Two smears were prepared from each individual; one smear was stained with one percent Aceto-orcein and the other with papanicolaou. Single blind method was used in which pathologist randomly analyzed 100 cells in each sample under oil immersion for identification of Barr bodies.For statistical analysis T-test was applied to compare the variance between aceto-orcein and PAP.The statistical significance was < P 0.05 RESULTS: The number of identified Barr bodies with aceto-orcein staining was significantly more when compared to Papanicolaou staining at P>0.05. CONCLUSION: Aceto-orcein proved to be a better stain to visualize nuclear details providing a higher accuracy in gender detection with remarkably less staining time than papanicolaou stain.