Department of Chemistry, Faculty of Science and Technology, Suan Sunandha Rajabhat University 1 U-thong, Nok Street, Wachira, Dusit, Bangkok 10300, Thailand.
Analyst. 2020 Jul 7;145(13):4637-4645. doi: 10.1039/d0an00515k. Epub 2020 May 27.
We report for the first time a highly sensitive and rapid quantitative method for the detection of Salmonella Typhimurium (S. Typhimurium) using a conductive immunosensor on a paper-based device (PAD). S. Typhimurium monoclonal antibodies (MA) were first immobilized on a paper-based device and then captured by S. Typhimurium. After an immunoreaction on the device, the polyclonal antibody-colloidal gold conjugate (PA-AuNPs) was dropped to bind with S. Typhimurium. After a complete sandwich reaction, a dark red color appeared on the paper-based device, which can be observed by the naked eye for a rapid screening test. The electrical conductivity of PA-AuNPs between the screen-printed electrodes on the paper-based device was also measured for an accurate quantitative analysis. The electrical conductivity correlated well with the concentration of S. Typhimurium, which was controlled by the amount of S. Typhimurium attached to the polyclonal antibody-colloidal gold conjugate. The device showed a linear correlation for the concentration of the S. Typhimurium in the range of 10-10 CFU mL in a logarithmic plot, with an R value of 0.9882 and a limit of detection (LOD) as low as 10 CFU mL. This simple, highly sensitive, and rapid method for the S. Typhimurium detection was successfully performed within 30 min, and it can be developed into small portable measuring devices in order to facilitate preliminary screening tests.
我们首次报道了一种基于纸基设备(PAD)的导电免疫传感器快速定量检测鼠伤寒沙门氏菌(S. Typhimurium)的高度敏感方法。首先将鼠伤寒沙门氏菌单克隆抗体(MA)固定在纸基设备上,然后捕获 S. Typhimurium。在设备上进行免疫反应后,将多克隆抗体-胶体金缀合物(PA-AuNPs)滴下以与 S. Typhimurium 结合。在完全三明治反应后,纸基设备上出现暗红色,可通过肉眼进行快速筛选测试。还测量了 PA-AuNPs 在纸基设备上的丝网印刷电极之间的电导率,以进行准确的定量分析。电导率与 S. Typhimurium 的浓度密切相关,该浓度受附着在多克隆抗体-胶体金缀合物上的 S. Typhimurium 数量控制。该设备在对数图中显示出 S. Typhimurium 浓度在 10-10 CFU mL 范围内的线性相关性,R 值为 0.9882,检测限(LOD)低至 10 CFU mL。这种简单、高度敏感和快速的 S. Typhimurium 检测方法可在 30 分钟内完成,并且可以开发成小型便携式测量设备,以方便进行初步筛选测试。