Norris Jessica V, Manning Kate, Linke Sarah J, Ferrance Jerome P, Landers James P
Department of Chemistry, McCormick Road, University of Virginia, Charlottesville, VA 22904, USA.
J Forensic Sci. 2007 Jul;52(4):800-5. doi: 10.1111/j.1556-4029.2007.00453.x. Epub 2007 May 25.
This report focuses on the development of a method for chemically induced enhancement of cell elution and recovery from cotton swabs. The method exploits the exclusive use of detergents for intact cell removal, and can be utilized in conjunction with, or to circumvent, conventional differential extraction (DE). Samples treated with Sarkosyl (54.4 +/- 1.8%) and sodium dodecyl sulfate (SDS) (78.5 +/- 0.7%) yielded higher sperm cell recoveries than a conventional DE buffer (39.4 +/- 2.1%). The results indicated that the choice of detergent affected sperm cell yield, with anionic detergents having the greatest effect. Storage time of samples affected the concentration of detergent required for optimal sperm cell recovery, longer times requiring increased detergent concentrations. In addition, the extent of sperm cell lysis by proteinase K digestion was evaluated. The results indicate that the exclusive use of SDS enhances the release of sperm and epithelial cells from a cotton swab as compared with DE buffer, providing for a more effective DNA analysis.
本报告重点介绍了一种通过化学诱导增强从棉签中洗脱和回收细胞的方法。该方法利用去污剂专门用于去除完整细胞，可与传统差异提取（DE）结合使用或规避其使用。用 Sarkosyl（54.4±1.8%）和十二烷基硫酸钠（SDS）（78.5±0.7%）处理的样本比传统DE缓冲液（39.4±2.1%）产生了更高的精子细胞回收率。结果表明，去污剂的选择会影响精子细胞产量，其中阴离子去污剂的影响最大。样本的储存时间会影响最佳精子细胞回收所需的去污剂浓度，储存时间越长，所需的去污剂浓度越高。此外，还评估了蛋白酶K消化导致的精子细胞裂解程度。结果表明，与DE缓冲液相比，单独使用SDS可增强从棉签中释放精子和上皮细胞的能力，从而实现更有效的DNA分析。
