警犬性控精液X、Y精子比例检测方法的建立OA
Establishment of a method to detect the ratio of X and Y sperm for sex-sorted semen in police dogs
为建立一种经济、快捷且准确的警犬精液X、Y精子比例检测方法,研究针对犬AKAP4(X精子标记)和SRY基因(Y精子标记)设计特异性引物,构建了系列已知拷贝数的质粒标准品;经实时荧光定量PCR,分别建立了AKAP4、SRY基因的标准曲线;通过检测混合质粒样本、市售性控精液及未分选精液对该方法进行验证.结果显示,AKAP4基因的标准曲线为y=-3.226x+40.412,决定系数(R²)=0.993,效率104%;SRY基因的标准曲线为y=-3.17x+37.225,R²=0.998,效率111%,线性关系良好.在不同稀释度和混合比例下,Y精子检测值与理论值差异均不显著(P>0.05),组间变异系数(CV)介于1.48%～2.73%之间,表明方法稳定可靠.研究表明,试验成功建立了一种操作简便、结果准确的警犬精液X、Y精子比例检测方法,可用于性控精液质量评估.
To establish an economical,rapid,and accurate method for detecting the proportion of X-and Y-chromosome-bearing sperm in police dog semen,specific primers were designed targeting canine AKAP4(an X-sperm marker)and SRY gene(a Y-sperm marker).A series of plasmid standards with known copy numbers were constructed.Standard curves for the AKAP4 and SRY genes were established by real-time quantitative PCR.The method was validated by detecting mixed plasmid samples,commercially available sex-sorted semen,and unsorted semen.The results showed that the standard curve for the AKAP4 gene was y=-3.226x+40.412,with an determination coefficient(R²)of 0.993 and an efficiency of 104%.The standard curve for the SRY gene was y=-3.17x+37.225,with an R² of 0.998 and an efficiency of 111%,indicating good linearity.At different dilution ratios and mixing proportions,there were no significant differences between the detected Y-sperm values and the theoretical values(P>0.05),and the coefficients of variation(CV)between groups ranged from 1.48%to 2.73%,demonstrating that the method is stable and reliable.This study successfully established a simple and accurate method for detecting the X/Y sperm ratio in police dog semen,which can be used for quality assessment of sex-sorted semen.
潘彩霞
中国刑事警察学院警犬技术学院,辽宁 沈阳 110000
农业科技
警犬性控精液精子比例荧光定量PCR
Police dogSex-sorted semenSperm ratioReal-time fluorescence quantitative PCR
《现代畜牧兽医》 2026 (4)
24-28,5
2025年度辽宁省教育厅高校基本科研项目(LJ212510175016)
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