Food, Nutrition and Health Program, Faculty of Land and Food Systems, The University of British Columbia, Vancouver, British Columbia, Canada.
Food, Nutrition and Health Program, Faculty of Land and Food Systems, The University of British Columbia, Vancouver, British Columbia, Canada
Appl Environ Microbiol. 2020 Apr 17;86(9). doi: 10.1128/AEM.00096-20.
spp. have been recognized as major foodborne pathogens worldwide. An increasing frequency of antibiotic-resistant pathogens, including spp., have been identified to transmit from food products to humans and cause severe threats to public health. To better mitigate the antibiotic resistance crisis, rapid detection methods are required to provide timely antimicrobial resistance surveillance data for agri-food systems. Herein, we developed a polymer-based microfluidic device for the identification and antimicrobial susceptibility testing (AST) of spp. An array of bacterial incubation chambers were created in the microfluidic device, where chromogenic medium and antibiotics were loaded. The growth of spp. was visualized by color change due to chromogenic reactions. This platform achieved 100% specificity for identification. Sensitive detection of multiple species (, , and ) was obtained in artificially contaminated milk and poultry meat, with detection limits down to 1 × 10 CFU/ml and 1 × 10 CFU/25 g, respectively. On-chip AST determined antibiotic susceptibilities by the lowest concentration of antibiotics that can inhibit bacterial growth (i.e., no color change observed). High coincidences (91% to 100%) of on-chip AST and the conventional agar dilution method were achieved against several clinically important antibiotics. For a presumptive colony, on-chip identification and AST were completed in parallel within 24 h, whereas standard methods, including biochemical assays and traditional culture-based AST, take several days for multiple sequential steps. In conclusion, this lab-on-a-chip device can achieve rapid and reliable detection of antibiotic-resistant spp. Increasing concerns of antibiotic-resistant spp. with regard to public health emphasize the importance of efficient and fast detection. This study described the timely identification and antimicrobial susceptibility testing of spp. by using a microfluidic device. Our developed method not only reduced the total analysis time, but it also simplified food sample preparation and chip operation for end users. Due to the miniaturized size of the lab-on-a-chip platform, the detection was achieved by using up to 1,000 times less of the reagents than with standard reference methods, making it a competitive approach for rapid screening and surveillance study in food industries. In addition, multiple clinically important species (, , and ) could be tested by our device. This device has potential for wide application in food safety management and clinical diagnostics, especially in resource-limited regions.
spp. 已被公认为全球主要的食源性病原体。越来越多的抗生素耐药病原体，包括 spp.，已被确定可从食品传播给人类，并对公众健康造成严重威胁。为了更好地缓解抗生素耐药危机，需要快速检测方法为农业食品系统提供及时的抗生素耐药监测数据。在此，我们开发了一种基于聚合物的微流控装置，用于 spp. 的鉴定和抗菌药敏试验 (AST)。在微流控装置中创建了一系列细菌孵育室，其中加载了显色培养基和抗生素。由于显色反应， spp. 的生长通过颜色变化可视化。该平台对 spp. 的鉴定具有 100%的特异性。在人工污染的牛奶和禽肉中，可检测到多种 spp.（ 、 、 ），检测限分别低至 1×10 CFU/ml 和 1×10 CFU/25 g。芯片上的 AST 通过可以抑制细菌生长的最低抗生素浓度（即观察到无颜色变化）来确定抗生素敏感性。针对几种临床重要抗生素，芯片上 AST 与传统琼脂稀释法的高度吻合率（91%至 100%）。对于假定的菌落，芯片上的鉴定和 AST 可在 24 小时内并行完成，而标准方法，包括生化检测和传统基于培养的 AST，需要数天时间进行多个连续步骤。总之，该芯片实验室装置可以实现快速可靠地检测抗生素耐药 spp. 鉴于公众健康对 spp. 抗生素耐药性的日益关注，强调了高效快速检测的重要性。本研究通过微流控装置描述了 spp. 的快速鉴定和抗菌药敏试验。我们开发的方法不仅缩短了总分析时间，而且简化了食品样品制备和芯片操作，方便了终端用户。由于该芯片实验室平台的小型化，与标准参考方法相比，检测所需的试剂用量减少了 1000 倍以上，使其成为食品工业中快速筛选和监测研究的一种有竞争力的方法。此外，我们的装置可以测试多种临床重要的 spp.（ 、 、 ）。该装置具有广泛应用于食品安全管理和临床诊断的潜力，特别是在资源有限的地区。
