Voorhees Jessica C, Ferrance Jerome P, Landers James P
Department of Chemistry, McCormick Road, University of Virginia, Charlottesville, VA 22904, USA.
J Forensic Sci. 2006 May;51(3):574-9. doi: 10.1111/j.1556-4029.2006.00112.x.
This report describes development of a method for enhanced cell elution from cotton swabs. The method exploits an enzyme mixture for digestion of the cotton to remove intact cells, and can be utilized in conjunction with or to circumvent conventional differential extraction (DE). Samples digested with Aspergillus niger cellulase yielded sperm cell recoveries (18+/-3.5%) similar to conventional DE buffer (23+/-7.8%) while providing intact epithelial cells. Storage time affected the concentration of enzyme required for optimal sperm cell recovery, with longer times requiring increased cellulase concentrations. Cellulase from A. niger yielded a twofold enhancement in sperm cell elution over buffer alone, and preliminary testing of higher activity cellulases from Trichoderma reesei and Trichoderma viride showed even greater enhancement. These results indicate that cellulose-digesting enzymes enhance the release of sperm and epithelial cells from a cotton swab over buffer alone, providing for efficient DNA analysis.
本报告描述了一种用于增强从棉签洗脱细胞的方法的开发。该方法利用一种酶混合物消化棉花以去除完整细胞，并且可以与传统的差异提取（DE）结合使用或用于规避传统的差异提取。用黑曲霉纤维素酶消化的样本产生的精子细胞回收率（18±3.5%）与传统的DE缓冲液（23±7.8%）相似，同时还能提供完整的上皮细胞。储存时间影响最佳精子细胞回收率所需的酶浓度，储存时间越长，所需的纤维素酶浓度越高。黑曲霉的纤维素酶比单独使用缓冲液能使精子细胞洗脱量提高两倍，对里氏木霉和绿色木霉更高活性纤维素酶的初步测试显示出更大的提高。这些结果表明，与单独使用缓冲液相比，纤维素消化酶能增强精子和上皮细胞从棉签上的释放，从而实现高效的DNA分析。
