The State Key Laboratory of Chemical Oncogenomics, Key Laboratory of Chemical Biology, Tsinghua Shenzhen International Graduate School, Tsinghua University, Shenzhen, 518055, China.
Shenzhen Lvshiyuan Biotechnology Co. Ltd, Shenzhen, 518120, China.
Biosens Bioelectron. 2022 Sep 15;212:114390. doi: 10.1016/j.bios.2022.114390. Epub 2022 May 17.
E. coli O157:H7 has become a crucial foodborne pathogen with certain morbidity and mortality. However, the available methods still face the significant challenges including the improvement of detection specificity and sensitivity for real sample analysis. Herein, a microfluidic chemiluminescence biosensor based on a multiple signal amplification strategy was developed for rapid and ultrasensitive detection of E.coli O157:H7. The microfluidic device was consisted of an upstream snake-shape mixing channel and a downstream ship-shape microcavity with micropillar array for high efficient mixing of CL reagents, sufficient CHA amplification reaction and CL reaction. The multiple signal amplification was achieved by bacteria competitively bind to trigger catalytic hairpin assembly (CHA) and HRP-Au NPs catalyzed CL reaction. Based on the platform, the detection of target bacteria is converted to the detection of nucleic acid with a limit detection down to 130 CFU/mL, which is better or comparable to previously reported biosensors. Moreover, the whole analysis time was about 1.5 h with only 10 μL of sample. The biosensor exhibited an excellent recovery rate ranged from 90.0% to 110.0% and good selectivity to the target bacteria. Overall, this developed biosensor is a promising tool for ultrahigh sensitive and rapid detection of foodborne pathogens.
产志贺毒素大肠杆菌 O157:H7 已成为一种重要的食源性致病菌，具有一定的发病率和死亡率。然而，现有的方法仍然面临着重大挑战，包括提高对实际样品分析的检测特异性和灵敏度。本文基于多重信号放大策略，开发了一种用于快速灵敏检测产志贺毒素大肠杆菌 O157:H7 的微流控化学发光生物传感器。微流控装置由上游蛇形混合通道和下游船形微腔以及微柱阵列组成，用于 CL 试剂的高效混合、充足的 CHA 放大反应和 CL 反应。通过细菌竞争结合触发催化发夹组装（CHA）和 HRP-Au NPs 催化 CL 反应实现了多重信号放大。基于该平台，目标细菌的检测转化为核酸的检测，检测限低至 130 CFU/mL，优于或可与先前报道的生物传感器相媲美。此外，整个分析时间约为 1.5 小时，仅需 10 μL 的样品。该生物传感器对目标细菌的回收率在 90.0%至 110.0%之间，表现出良好的回收率和良好的选择性。总的来说，这种开发的生物传感器是一种用于超灵敏快速检测食源性致病菌的有前途的工具。
