Chemistry and Biochemistry Department, University of Maryland, Baltimore County, Baltimore, MD, United States of America.
Institute of Fluorescence, University of Maryland, Baltimore County, Baltimore, MD, United States of America.
PLoS One. 2019 Jul 23;14(7):e0220102. doi: 10.1371/journal.pone.0220102. eCollection 2019.
The ability for safe and rapid pathogenic sample transportation and subsequent detection is an increasing challenge throughout the world. Herein, we describe and use bead-beating, vortex, sonication, 903 protein saver cards, and Lyse-It methods, aiming to inactivate Gram-positive and -negative bacteria with subsequent genome DNA (quantitative Polymerase Chain Reaction) qPCR detection. The basic concepts behind the four chosen technologies is their versatility, cost, and ease of use in developed and underdeveloped countries. The four methods target the testing of bacterial resilience, cellular extraction from general and complex media and subsequent DNA extraction for qPCR detection and amplification. These results demonstrate that conventional high temperature heating, 903 protein saver cards, and Lyse-It are all viable options for inactivating bacterial growth for safe shipping. Additionally, Lyse-It was found to be particularly useful as this technology can inactivate bacteria, extract cells from 903 protein saver cards, lyse bacterial cells, and additionally keep genomic DNA viable for qPCR detection.
安全快速地进行病原体样本运输和后续检测的能力是全世界日益面临的挑战。在此，我们描述并使用了珠磨、涡旋、超声、903 蛋白保存卡和 Lyse-It 方法，旨在灭活革兰氏阳性和阴性细菌，随后进行基因组 DNA（定量聚合酶链反应）qPCR 检测。这四种技术背后的基本概念是它们的多功能性、成本效益以及在发达国家和发展中国家的易用性。这四种方法旨在测试细菌的抗逆性、从一般和复杂介质中提取细胞，以及随后进行 qPCR 检测和扩增的 DNA 提取。这些结果表明，常规高温加热、903 蛋白保存卡和 Lyse-It 都是安全运输中灭活细菌生长的可行选择。此外，Lyse-It 被发现特别有用，因为这项技术可以灭活细菌，从 903 蛋白保存卡中提取细胞，裂解细菌细胞，并保持基因组 DNA 用于 qPCR 检测的活性。
