Dept. of Chemical Engineering, Indian Institute of Technology (IIT) Delhi, 110016, India.
Molecular Diagnostics and Biomarkers Lab, Gleneagles Global Hospitals, Lakdi ka pul, Hyderabad, 500004, India.
Biosens Bioelectron. 2022 Mar 15;200:113876. doi: 10.1016/j.bios.2021.113876. Epub 2021 Dec 18.
The present antibiotic susceptibility testing (AST) techniques based on bacterial culture, gene amplification and mass spectrometry are highly time consuming, labour intensive or expensive. Impedance spectroscopy is an emerging tool for rapid bacterial analysis as it is label-free, real-time, affordable and high-throughput. The over-reliance of this technique on complex chip designs and cell enrichment strategies has, however, slowed its foray into clinical AST. We demonstrate a label-free approach in which a low conductivity zwitterionic buffer is used for boosting impedance sensitivity in simple interdigitated electrodes (IDEs) allowing rapid AST in just 20 min without any liquid flow, biofunctionalization or cell enrichment steps. The detection principle relies on measuring changes in solution resistance due to antibiotic-induced bacterial cell death or growth. While the death-based approach is faster (20 min), it's restricted to surface-acting bactericidal antibiotics. The cell growth approach is longer (60-80 min) but more versatile as it applies to all drug types. Results for antibiotic sensitivity analysis and minimum inhibitory concentration (MIC) determination are illustrated for Escherichia coli, Klebsiella pneumoniae and Staphylococcus aureus against a wide class of antibiotics (penicillins, cephalosporins, polymyxins, carbapenems etc.).
目前基于细菌培养、基因扩增和质谱的抗生素药敏检测(AST)技术耗时、劳动强度大或成本高。阻抗谱分析是一种新兴的快速细菌分析工具,因为它是无标记的、实时的、经济实惠的和高通量的。然而,这种技术对复杂的芯片设计和细胞富集策略的过度依赖,已经减缓了它在临床 AST 中的应用。我们展示了一种无标记的方法,其中使用低电导率两性离子缓冲液来提高简单叉指电极(IDE)的阻抗灵敏度,从而在没有任何液体流动、生物功能化或细胞富集步骤的情况下仅用 20 分钟即可实现快速 AST。检测原理依赖于测量由于抗生素诱导的细菌细胞死亡或生长而导致的溶液电阻的变化。虽然基于死亡的方法更快(20 分钟),但它仅限于表面作用的杀菌抗生素。细胞生长方法耗时更长(60-80 分钟),但适用性更广,适用于所有药物类型。针对广泛类别的抗生素(青霉素、头孢菌素、多粘菌素、碳青霉烯等),我们对大肠杆菌、肺炎克雷伯菌和金黄色葡萄球菌进行了抗生素敏感性分析和最小抑菌浓度(MIC)测定的结果。