Leonard R B, Carroll K C
Associated Regional and University Pathologists, Inc., Salt Lake City, Utah, USA.
Diagn Mol Pathol. 1997 Oct;6(5):288-91. doi: 10.1097/00019606-199710000-00007.
To adapt pulsed-field gel electrophoresis (PFGE) to a busy clinical laboratory, we evaluated the use of achromopeptidase (ACP) as the sole agent for lysis of a variety of gram-positive cocci. Growth in an appropriate broth media was adjusted to a 3.0 McFarland density before aliquotting for washing and plug preparation. ACP incorporated into the agarose plugs, or added to the buffer solution, lysed cells in < 1 h when suspended in a Tris-NaCl buffer. Two 30-min washes in a Tris-EDTA buffer were sufficient to remove cellular debris without additional deproteination. Final PFGE results were obtained in approximately 48 h, including the initial broth subculturing.
为使脉冲场凝胶电泳（PFGE）适用于繁忙的临床实验室，我们评估了使用无色肽酶（ACP）作为裂解多种革兰氏阳性球菌的唯一试剂。在分装用于洗涤和制备琼脂糖块之前，将其在合适的肉汤培养基中的生长调整至3.0麦氏浊度。当悬浮于Tris - NaCl缓冲液中时，掺入琼脂糖块或添加到缓冲溶液中的ACP可在1小时内裂解细胞。在Tris - EDTA缓冲液中进行两次30分钟的洗涤足以去除细胞碎片，无需额外的脱蛋白处理。最终的PFGE结果在约48小时内获得，包括最初的肉汤传代培养。
