A simple method for gender verification based on PCR detection of Y-chromosomal DNA and its application at the Winter Universiade 1991 in Sapporo City, Japan.

Abstract

A simple and reliable screening method for gender verification at international sports competitions was developed on the basis of detection of Y-chromosomal DNA which was specifically amplified by polymerase chain reaction (PCR). Total DNA obtained from buccal mucous membrane cells by digestion with proteinase K followed by boiling treatment was directly used for the PCR. The amplified product was electrophoresed on an agarose gel staining with ethidium bromide. We adopted three sets of the PCR primer, identifying different regions of Y-chromosome, of which sensitivity and specificity were preliminary evaluated for 228 (112 male, 116 female) DNA samples from lymphocytes. This new method was first applied for the gender verification test at the Winter Universiade 1991 Sapporo, accompanied by traditional microscopic tests for X- and Y-chromatins. No positive results were obtained for 155 female competitors using both the PCR and the microscopic methods. The superiority of the proposed method was clearly shown in the reliability of the results and also in the saving on instrumental and personnel costs as compared to previous cytologic methods.