Rapid detection and identification of bacteria: SEnsing of phage-triggeredion cascade (SEPTIC)
Rapid detection and identification of bacteria: SEnsing of phage-triggeredion cascade (SEPTIC)
复制标题
DOI：
10.4024/1050501.jbpc.05.01
复制
发表时间：
2005-01-01
期刊：
影响因子：
--
通讯作者：
Kish, L. B.
中科院分区：
文献类型：
--
作者：
Dobozi-King, M.;Seo, S.;Kish, L. B.
因此，迫切需要快速检测和鉴定细菌的方法。在这里，我们描述了一种方法，结合噬菌体的特异性和亲和力与电噪声的波动分析。该方法是基于在噬菌体DNA注入宿主细胞的瞬间发生的大量瞬时离子泄漏。离子通量仅需要细胞是生理上可存活的（即具有通电的膜），并且可以在将细胞与足够浓度的噬菌体颗粒混合后的几秒钟内发生。为了检测这些通量，我们构建了一个纳米井，一个横向的，微米大小的钛电极电容器，间隙大小为150 nm，并使用它来测量含有噬菌体和细菌的微升（mm（3））样品中的电场波动。在分析物细菌对噬菌体敏感的混合物中，观察到具有各种时间和振幅尺度的大随机波，功率谱近似遵循1/f（2）定律，从1-10 Hz。这种SEPTIC（SEensing of Phage-Triggered Ion Cascades）技术的开发可以在几分钟内快速检测和鉴定活病原菌，具有无与伦比的特异性。该方法的潜在最终灵敏度为1个细菌/微升（1个细菌/mm（3））。
Methods for the rapid detection and identification of bacteria are urgently needed. Here we describe a method that combines the specificity and avidity of bacteriophages with fluctuation analysis of electrical noise. The method is based on the massive transitory ion leakage that occurs at the moment of phage DNA injection into the host cell. The ion fluxes require only that the cells be physiologically viable (i.e. have energized membranes) and can occur within seconds after mixing the cells with sufficient concentrations of phage particles. To detect these fluxes, we have constructed a nano-well, a lateral, micrometer-sized capacitor of titanium electrodes with gap size of 150 nm, and used it to measure the electrical field fluctuations in microlitre (mm(3)) samples containing phage and bacteria. In mixtures where the analyte bacteria were sensitive to the phage, large stochastic waves with various time and amplitude scales were observed, with power spectra approximately following a 1/f(2) law from 1-10 Hz. Development of this SEPTIC (SEnsing of Phage-Triggered Ion Cascades) technology could provide rapid detection and identification of live pathogenic bacteria on the scale of minutes, with unparalleled specificity. The method has a potential ultimate sensitivity of 1 bacterium/microlitre (1 bacterium/mm(3)).