Yamaguchi A, Fukushi M, Kikuchi Y, Aparicio J, Wakisaka A
Sapporo City Institute of Public Health, Japan.
Int J Sports Med. 1992 May;13(4):304-7. doi: 10.1055/s-2007-1021271.
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.
基于聚合酶链反应（PCR）特异性扩增的Y染色体DNA检测，开发了一种简单可靠的国际体育比赛性别验证筛查方法。用蛋白酶K消化颊粘膜细胞，随后进行煮沸处理，从中获得的总DNA直接用于PCR。扩增产物在含有溴化乙锭的琼脂糖凝胶上进行电泳。我们采用了三组PCR引物，用于识别Y染色体的不同区域，对来自淋巴细胞的228份（112份男性，116份女性）DNA样本初步评估了其敏感性和特异性。这种新方法首次应用于1991年札幌冬季大学生运动会的性别验证测试，同时还采用了传统的X和Y染色质显微镜检测方法。使用PCR和显微镜方法对155名女性参赛者均未得到阳性结果。与之前的细胞学方法相比，该方法在结果可靠性以及节省仪器和人员成本方面明显显示出优势。
