显微操作法提取混合斑中精子细胞方法的探讨

The exploring of a method for isolating sperm cells from mixture sample

目的尝试建立一种提取混合斑中精子细胞的检测方法。方法在人为控制条件下,制备精液—阴道液混合斑,分别使用显微操作法与差异裂解法分离精子细胞,提取DNA,进行STR基因型检测。结果采用显微操作法检测成功率11/12,差异裂解法成功率1/12,两者有显著性差异。结论显微操作法可有效获取精子细胞,排除女性物质和其它杂质的干扰,STR分型成功率优于差异裂解法。

Objective To establish a method for isolation of sperm cells from mixture sample by micromanipulation method, in order to improve the performance of STR typing in those cases where male DNA availability is limited. Methods Mixture samples were prepared with sperm cells and vaginal swabs under controlled conditions. Both the micromanipulation method and the differential lysis method were employed to obtain sperm cells from the mixture samples. STR genotyping was carried out with the obtained sperm cells. Results The micromanipulation method with genotyping success in 11 out of 12 samples was found to outperform the differential lysis method, while sample from the differential lysis method was genotyped only in 1 out of 12 samples. Difference between the two results was significant. Conclusion This study showed micromanipulation method was efficient to the isolation of sperm cells from mixture sample and was enough to meet the need of STR genotyping of sperm cells.