Wu Yutong, Liu Yang, Ma Jinchen, Zhu Shanxi, Zhao Xiaojun, Mou Huawei, Ke Xuanqi, Wu Zhisheng, Wang Yifei, Lin Sheng, Qi Wuzhen
School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China.
School of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan 250355, China.
Biosensors (Basel). 2024 Dec 27;15(1):10. doi: 10.3390/bios15010010.
Microbial contamination is an important factor threatening the safety of Chinese medicine preparations, and microfluidic detection methods have demonstrated excellent advantages in the application of rapid bacterial detection. In our study, a novel optical biosensor was developed for the rapid and sensitive detection of in traditional Chinese medicine on a microfluidic chip. Immune gold@platinum nanocatalysts (Au@PtNCs) were utilized for specific bacterial labeling, while magnetic nano-beads (MNBs) with a novel high-gradient magnetic field were employed for the specific capture of bacteria. The immune MNBs, immune Au@PtNCs, and bacterial samples were introduced into a novel passive microfluidic micromixer for full mixing, resulting in the formation of a double-antibody sandwich structure due to antigen-antibody immune reactions. Subsequently, the mixture flowed into the reaction cell, where the MNBs--Au@PtNCs complex was captured by the magnetic field. After washing, hydrogen peroxide-tetramethylbenzidine substrate (HO-TMB) was added, reacting with the Au@PtNCs peroxidase to produce a blue reaction product. This entire process was automated using a portable device, and concentration was analyzed via a phone application. This simple biosensor has good specificity with a detection range of 9 × 10-9 × 10 CFU/mL and can detect concentrations as low as 90 CFU/mL within 74 min. The average recoveries of the spiked samples ranged from 76.8% to 109.5.
微生物污染是威胁中药制剂安全性的重要因素，而微流控检测方法在快速细菌检测应用中已展现出卓越优势。在我们的研究中，开发了一种新型光学生物传感器，用于在微流控芯片上对中药中的[具体细菌名称未给出]进行快速灵敏检测。免疫金@铂纳米催化剂（Au@PtNCs）用于特异性细菌标记，而具有新型高梯度磁场的磁性纳米珠（MNBs）用于细菌的特异性捕获。将免疫MNBs、免疫Au@PtNCs和细菌样品引入新型被动微流控微混合器进行充分混合，由于抗原 - 抗体免疫反应形成双抗体夹心结构。随后，混合物流入反应池，在那里MNBs - Au@PtNCs复合物被磁场捕获。洗涤后，加入过氧化氢 - 四甲基联苯胺底物（HO - TMB），与Au@PtNCs过氧化物酶反应产生蓝色反应产物。整个过程使用便携式设备自动化进行，[具体细菌名称未给出]浓度通过手机应用程序进行分析。这种简单的生物传感器具有良好的特异性，检测范围为9×10 - 9×10 CFU/mL，并且能够在74分钟内检测低至90 CFU/mL的[具体细菌名称未给出]浓度。加标样品的平均回收率在76.8%至109.5%之间。
