Objective: Buccal epithelial cells in saliva traces found at a crime scene can be used for sex determination by examining the presence of Barr bodies in the nucleus. Papanicolaou (PAP) and fluorescent stains, among others, have been used in the past. Aceto-orcein (AO) is a relatively unexplored staining technique for this purpose. This study aims to assess the efficacy of sex determination using AO and PAP stains for the detection of Barr bodies in buccal mucosal scrapes. Study Design: Buccal scrapings were collected from 120 healthy individuals (60 males and 60 females). They were stained with AO and PAP. Fifty cells in each sample were analyzed for identification of Barr bodies. Samples with a presence of Barr bodies ≤5% were recorded as male and those with >5% were recorded as female. The percentage accuracy in determining sex using both stains was evaluated. Results: The percentage of Barr bodies in AO-stained slides ranged from 5 to 32 among females and from 0 to 8 in males, while with PAP the ranges recorded were 4-20 in females and 0-5 in males (p < 0.001). The sensitivity and specificity of AO for detecting sex accurately was around 98.3 and 95% for PAP. Conclusion: Sex determination using Barr bodies in the buccal cells is a simple method that provides up to 95-98% accuracy; making it a significant accessory for sex determination. AO proved to be a better stain than PAP for visualizing nuclear details, and its staining time was remarkably shorter. It also demonstrated enhanced sex estimation efficacy compared to PAP.

1.
Barr ML, Carr DH: Sex chromatin, sex chromosomes and sex anomalies. Can Med Assoc J 1960;83:979-986.
2.
Mittwoch U: Human and animal chromosomes: sex chromatin. Proc R Soc Med 1964;57:643-645.
3.
Sanderson AR, Stewart JS: Nuclear sexing with aceto-orcein. Br Med J 1961;2:1065-1067.
4.
Manjula Bhai KH, Yadwad BS, Patil PV: A study of Barr bodies in Indian, Malaysian and Chinese subjects. J Forensic Med Toxicol 1997;14:9-13.
5.
Platt LI, Kalin EW: Sex chromatin frequency. JAMA 1964;187:94-98.
6.
DeGraaf RM, Larson JS: Technique for the observation of sex chromatin in hair roots. J Mammal 1972 ;53:368-371.
7.
Suazo GI, Roa HI, Cantín LM: Sex chromatin in dental pulp: performance of diagnosis test and gold standard generation. Int J Morphol 2010;28:1093-1096.
8.
Nagamori H, Ohno Y, Uchima E, Kajiwara M, Nakazato M, Une Y, Takeda K: Sex determination from buccal mucosa and hair root by the combined treatment of quinacrine staining and the fluorescent Feulgen reaction using a single specimen. Forensic Sci Int 1986;31:119-128.
9.
Anoop UR, Ramesh V, Balamurali PD, Nirima O, Premalatha B, Karthikshree VP: Role of Barr bodies obtained from oral smears in the determination of sex. Indian J Dent Res 2004;15:5-7.
10.
Bancroft JD, Stevens A: Theories and Practice of Histological Techniques, ed 4. London, Churchill Livingstone, pp 114-150.
11.
Mittal T, Saralaya KM, Kuruvilla A, Archary C: Sex determination from buccal mucosa scrapes. Int J Legal Med 2009;123:437-440.
12.
Aggarwal NK, Kumar S, Banerjee KK, Agarwal BB: Sex determination from buccal mucosa. J Forensic Med Toxicol 1996;13:43-44.
13.
Dixon AD, Torr JB: Sex chromatin in oral smears. Br Med J 1956;2:799-800.
14.
Gardner HA: The buccal smear: reassessment of its usefulness. Can Med Assoc J 1976;114:527-530.
15.
Smith DW, Marden PM, McDonald MJ, Spechard M: Lower incidence of sex chromatin in buccal smears of newborn females. Pediatrics 1962;707:11-12.
16.
Gonzales HC, Mendes FT, Saldanha PH: Sex chromatin in normal newborns in first two weeks of life: a blind study. Rev Brasil Genet 1978;4:263-278.
17.
Hagy GW, Brodrick MM: Variation of sex chromatin in human oral mucosa during menstrual cycle. Acta Cytol 1972;16:314-320.
18.
Hamerton JL, Jagiello GM, Kirman BH: Sex-chromosome abnormalities in a population of mentally defective children. Br Med J 1962;1:220-223.
19.
Ghosh SN, Shah PN, Desai PB: Barr body frequency in esophageal cancer in Indian women. Acta Cytol 1983;27:202-203.
20.
Nishiya I, Ishizaki Y, Sasaki M: Nuclear DNA content and the number of Barr bodies in premalignant and malignant lesions of the uterine cervix. Acta Cytol 1981;25:407-411.
21.
Oliveria AM, French CA: Applications of fluorescence in situ hybridization in cytopathology. Acta Cytol 2005;49:587-594.
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.