Suppr超能文献

革兰氏阳性菌和革兰氏阴性菌的电化学细胞裂解:从环境水样中提取DNA

Electrochemical cell lysis of gram-positive and gram-negative bacteria: DNA extraction from environmental water samples.

作者信息

Wang Siwen, Zhu Yanzhe, Yang Yang, Li Jing, Hoffmann Michael R

机构信息

Linde+Robinson Laboratories, California Institute of Technology, Pasadena, CA 91125, USA.

出版信息

Electrochim Acta. 2020 Apr 1;338:135864. doi: 10.1016/j.electacta.2020.135864.

Abstract

Cell lysis is an essential step for the nucleic acid-based surveillance of bacteriological water quality. Recently, electrochemical cell lysis (ECL), which is based on the local generation of hydroxide at a cathode surface, has been reported to be a rapid and reagent-free method for cell lysis. Herein, we describe the development of a milliliter-output ECL device and its performance characterization with respect to the DNA extraction efficiency for gram-negative bacteria ( and Typhi) and gram-positive bacteria ( and ). Both gram-negative and gram-positive bacteria were successfully lysed within a short but optimal duration of 1 min at a low voltage of ∼5 V. The ECL method described herein, is demonstrated to be applicable to various environmental water sample types, including pond water, treated wastewater, and untreated wastewater with DNA extraction efficiencies similar to a commercial DNA extraction kit. The ECL system outperformed homogeneous chemical lysis in terms of reaction times and DNA extraction efficiencies, due in part to the high pH generated at the cathode surface, which was predicted by simulations of the hydroxide transport in the cathodic chamber. Our work indicates that the ECL method for DNA extraction is rapid, simplified and low-cost with no need for complex instrumentation. It has demonstrable potential as a prelude to PCR analyses of waterborne bacteria in the field, especially for the gram-negative ones.

摘要

细胞裂解是基于核酸的细菌水质监测的关键步骤。最近,据报道,基于阴极表面局部产生氢氧化物的电化学细胞裂解(ECL)是一种快速且无需试剂的细胞裂解方法。在此,我们描述了一种毫升级输出的ECL装置的开发及其对革兰氏阴性菌(伤寒杆菌和鼠伤寒沙门氏菌)和革兰氏阳性菌(金黄色葡萄球菌和枯草芽孢杆菌)DNA提取效率的性能表征。革兰氏阴性菌和革兰氏阳性菌均在约5 V的低电压下,于1分钟的短最佳持续时间内成功裂解。本文所述的ECL方法被证明适用于各种环境水样类型,包括池塘水、处理后的废水和未处理的废水,其DNA提取效率与商业DNA提取试剂盒相似。ECL系统在反应时间和DNA提取效率方面优于均相化学裂解,部分原因是阴极表面产生的高pH值,这是通过阴极室中氢氧化物传输的模拟预测的。我们的工作表明,用于DNA提取的ECL方法快速、简便且成本低,无需复杂的仪器。它作为现场对水生细菌进行PCR分析的前奏具有明显的潜力,特别是对于革兰氏阴性菌。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/33b3/7063685/be585c3a162a/fx1.jpg

文献检索

告别复杂PubMed语法,用中文像聊天一样搜索,搜遍4000万医学文献。AI智能推荐,让科研检索更轻松。

立即免费搜索

文件翻译

保留排版,准确专业,支持PDF/Word/PPT等文件格式,支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述,25分钟生成高质量综述,智能提取关键信息,辅助科研写作。

立即免费体验