Redox cycling-based detection of phenazine metabolites secreted from Pseudomonas aeruginosa in nanopore electrode arrays.
Redox cycling-based detection of phenazine metabolites secreted from Pseudomonas aeruginosa in nanopore electrode arrays.
复制标题
DOI：
10.1039/d0an02022b
复制
发表时间：
2021-02-21
期刊：
影响因子：
--
通讯作者：
Bohn PW
中科院分区：
文献类型：
--
作者：
Do H ;Kwon SR ;Baek S ;Madukoma CS ;Smiley MK ;Dietrich LE ;Shrout JD ;Bohn PW
机会致病菌铜绿假单胞菌（P. aeruginosa）产生几种氧化还原活性吩嗪代谢物，包括绿脓菌素（PYO）和吩嗪-1-甲酰胺（PCN），它们是电子载体分子，也有助于毒力。具体而言，PYO是由铜绿假单胞菌产生的唯一代谢物，其在医院获得性感染中充当毒力因子，因此是用于鉴定该病原体的早期定殖的良好生物标志物。在这里，我们描述了使用纳米孔电极阵列（NEAs）表现出金属-绝缘体-金属环电极架构，用于增强检测这些吩嗪代谢物。纳米孔的大小允许吩嗪代谢物自由扩散到内部并进入工作电极，同时排除细菌。因此，NEA中的高效氧化还原循环反应可以通过不受细菌存在阻碍的自由扩散进行。该策略产生低检测限，即PYO和PCN分别为10.5和20.7 nM，远低于单分子孔占有率的值，例如在10.5 nM孔占有率/纳米孔~ 0.082-反映NEA中异常信号放大的极限。此外，比较基本培养基和丰富培养基结果的实验表明，铜绿假单胞菌产生相同类型的吩嗪代谢物，即使在这两种培养基中生长速率和吩嗪产生模式不同。这里开发的NEA测量策略应该是有用的病原体一般的诊断和了解代谢在临床上重要的微生物群落。
The opportunistic pathogen Pseudomonas aeruginosa (P. aeruginosa) produces several redox-active phenazine metabolites, including pyocyanin (PYO) and phenazine-1-carboxamide (PCN), which are electron carrier molecules that also aid in virulence. In particular, PYO is an exclusive metabolite produced by P. aeruginosa, which acts as a virulence factor in hospital-acquired infections and is therefore a good biomarker for identifying early stage colonization by this pathogen. Here, we describe the use of nanopore electrode arrays (NEAs) exhibiting metal-insulator-metal ring electrode architectures for enhanced detection of these phenazine metabolites. The size of the nanopores allows phenazine metabolites to freely diffuse into the interior and access the working electrodes, while the bacteria are excluded. Consequently, highly efficient redox cycling reactions in the NEAs can be accessed by free diffusion unhindered by the presence of bacteria. This strategy yields low limits of detection, i.e. 10.5 and 20.7 nM for PYO and PCN, respectively, values far below single molecule pore occupancy, e.g. at 10.5 nM 〈npore〉 ~ 0.082 per nanopore - a limit which reflects the extraordinary signal amplification in the NEAs. Furthermore, experiments that compared results from minimal medium and rich medium show that P. aeruginosa produces the same types of phenazine metabolites even though growth rates and phenazine production patterns differ in these two media. The NEA measurement strategy developed here should be useful as a diagnostic for pathogens generally and for understanding metabolism in clinically important microbial communities.
登录
查看更多内容
影响因子：
4.6
作者：
Buzid A;Shang F;Reen FJ;Muimhneacháin EÓ;Clarke SL;Zhou L;Luong JH;O'Gara F;McGlacken GP;Glennon JD
通讯作者：
Glennon JD
影响因子：
3.6
作者：
Dietrich, Lars E. P.;Price-Whelan, Alexa;Newman, Dianne K.
通讯作者：
Newman, Dianne K.
影响因子：
2.6
作者：
ALANI, FY;ALSHIBIB, AS;TAHER, R
通讯作者：
TAHER, R
影响因子：
3.2
作者：
Mavrodi, DV;Bonsall, RF;Thomashow, LS
通讯作者：
Thomashow, LS
影响因子：
9.4
作者：
Chen, Jonathan H. K.;Ho, Pak-Leung;Yam, Wing-Cheong
通讯作者：
Yam, Wing-Cheong