Feng Ye, Zhou Dan, Gao Lujia, He Fengjiao
State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082, China.
Chemistry Department, Reed College, Portland, 97202, USA.
Biosens Bioelectron. 2020 Nov 15;168:112527. doi: 10.1016/j.bios.2020.112527. Epub 2020 Aug 22.
The early detection of bacteria is of critical importance in addressing serious public health problems. Here, an electrochemical biosensor for rapid detection of bacteria based on facile synthesis of silver wire across electrodes was constructed. High-variable region of 16S rRNA of bacteria was used as biomarker. Polymerase-free synthesis of silver wire was introduced into electrochemical signal transduction to improve the sensitivity of electrochemical detection. The construction biosensor of proposed method is as follows: Metastable hairpin probe H was modified on electrode surface, biomarker can open the stem-loop structure of H and activates HCR. The alternate opening of the stem-loop structure of H and H-AuNPs finally results in the formation of long double-stranded DNA-RNA (HCR products) -AuNPs. The formed AuNPs modified HCR products was blown in one direction using N to across the electrode gap. Using this HCR products as template, the silver wire was formed between the electrodes by silver deposition, and resulted in sharp change in electrical parameters of electrode. As the proof-of-concept work, multichannel series piezoelectric quartz crystal (MSPQC) was utilized as detector. The detection of Staphylococcus aureus in the concentration range from 50 to 10 CFU/mL within 100 min was achieved. The detection limit was 50 CFU/mL. Escherichia coli, Salmonella enteritidis, Listeria innocua, Pseudomonas aeruginosa and Streptococcus pneumoniae did not interfere the detection results. This newly proposed electrochemical biosensor is simple, rapid and exhibit high signal-to-noise ratio, it has great potential for being applied in food safety monitoring and clinical diagnosis.
细菌的早期检测对于解决严重的公共卫生问题至关重要。在此,构建了一种基于在电极上简便合成银线的用于快速检测细菌的电化学生物传感器。细菌16S rRNA的高变区用作生物标志物。将无聚合酶的银线合成引入电化学信号转导以提高电化学检测的灵敏度。所提出方法的生物传感器构建如下:亚稳态发夹探针H修饰在电极表面,生物标志物可打开H的茎环结构并激活HCR。H和H-AuNPs的茎环结构交替打开最终导致形成长双链DNA-RNA(HCR产物)-AuNPs。使用N将形成的修饰有AuNPs的HCR产物吹向一个方向以穿过电极间隙。以该HCR产物为模板,通过银沉积在电极之间形成银线,并导致电极电参数的急剧变化。作为概念验证工作,利用多通道串联压电石英晶体(MSPQC)作为检测器。在100分钟内实现了对金黄色葡萄球菌在50至10 CFU/mL浓度范围内的检测。检测限为50 CFU/mL。大肠杆菌、肠炎沙门氏菌、无害李斯特菌、铜绿假单胞菌和肺炎链球菌不干扰检测结果。这种新提出的电化学生物传感器简单、快速且具有高信噪比,在食品安全监测和临床诊断中具有巨大的应用潜力。