State Key Laboratory of Food Science and Technology, School of Food Science and Technology, National Engineering Research Center for Functional Food, Synergetic Innovation Center of Food Safety and Quality Control, Jiangnan University, Wuxi, 214122, Jiangsu, China.
State Key Laboratory of Dairy Biotechnology,Shanghai Engineering Research Center of Dairy Biotechnology, Dairy Research Institute, Bright Dairy & Food Co., Ltd., Shanghai, 200436, Shanghai, China.
Anal Bioanal Chem. 2021 Feb;413(4):1127-1136. doi: 10.1007/s00216-020-03076-8. Epub 2021 Jan 8.
Antibiotic resistance has become a serious threat to food safety and public health globally. Therefore, the development of a sensitive, quick, and simple method for antibiotic susceptibility testing is an urgent and crucial need. A novel concentration gradient microfluidic chip was designed in this work to generate antibiotic concentration gradient, culture bacteria, and produce fluorescence emission. An in-house-assembled fluorescence detection platform was constructed, and experiments were conducted to verify the linearity of the generated concentration gradient, explore the appropriate incubation time and flow rate for the microfluidic chip, and study the effect of long-term acid-based food processing on antibiotic susceptibility testing. Experimental results show that the concentration gradient generated by the microfluidic chip exhibited good linearity, stability, and controllability. The appropriate flow rate and incubation time for the microfluidic chip were 2 μL/min and 5 h, respectively. The use of this microfluidic chip for testing antibiotic resistance of Salmonella to ofloxacin and ampicillin generated results that were completely consistent with test results obtained using the gold-standard method. Furthermore, Salmonella showed greater sensitivity to antibiotics under strong acid conditions, confirming the potential influence of acid-based food processing on antibiotic susceptibility testing of real samples. The designed microfluidic chip provides a high-throughput, sensitive, and rapid antibiotic susceptibility testing method that combines the microfluidic chip and the fluorescence detection platform. The application of this method would facilitate determination of antibiotic-resistant bacterial strains for clinicians and researchers, and enable monitoring of changes in bacterial resistance during food processing.
抗生素耐药性已成为全球食品安全和公共卫生的严重威胁。因此，开发一种灵敏、快速、简单的抗生素药敏检测方法是当务之急。本工作设计了一种新型浓度梯度微流控芯片，用于生成抗生素浓度梯度、培养细菌并产生荧光发射。构建了一个内部组装的荧光检测平台，并进行了实验以验证所生成浓度梯度的线性度，探索微流控芯片的合适孵育时间和流速，以及研究长期酸性食品加工对抗生素药敏检测的影响。实验结果表明，微流控芯片生成的浓度梯度具有良好的线性度、稳定性和可控性。微流控芯片的合适流速和孵育时间分别为 2 μL/min 和 5 h。使用该微流控芯片对沙门氏菌对氧氟沙星和氨苄西林的抗生素耐药性进行测试，得到的结果与金标准方法完全一致。此外，在强酸条件下，沙门氏菌对抗生素表现出更高的敏感性，证实了酸性食品加工对实际样品抗生素药敏检测的潜在影响。设计的微流控芯片提供了一种高通量、灵敏、快速的抗生素药敏检测方法，将微流控芯片和荧光检测平台结合在一起。该方法的应用将有助于临床医生和研究人员确定耐药菌菌株，并监测食品加工过程中细菌耐药性的变化。
