2024-07-18
【學術亮點】免標定式介電泳分選平台用於細菌抗藥性之評估
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【學術亮點】免標定式介電泳分選平台用於細菌抗藥性之評估
Intelligent Husbandry: AI Green Energy Management and Circular Use of Sensing Platform【Department of Bio-industrial Mechatronics Engineering/ Ching-Chou Wu / Professor】
智慧農牧場：AI綠能管理與循環利用之感測平台【生機系吳靖宙教授】
上架日期：2024/4/26
Intelligent Husbandry: AI Green Energy Management and Circular Use of Sensing Platform【Department of Bio-industrial Mechatronics Engineering/ Ching-Chou Wu / Professor】
智慧農牧場：AI綠能管理與循環利用之感測平台【生機系吳靖宙教授】
| 論文篇名 | 英文：A Label-Free Droplet Sorting Platform Integrating Dielectrophoretic Separation for Estimating Bacterial Antimicrobial Resistance 中文：免標定式介電泳分選平台用於細菌抗藥性之評估 |
| 期刊名稱 | Biosensors |
| 發表年份, 卷數, 起迄頁數 | 2024, 14, 218 |
| 作者 | Jia-De Yan, Chiou-Ying Yang, Arum Han (美國學者), Ching-Chou Wu(吳靖宙)* |
| DOI | 10.3390/bios14050218 |
| 中文摘要 | 抗微生物耐藥性（AMR）已成為全球健康問題。抗生素耐藥細菌在抗生素治療後仍可存活，會降低療效並增加致命風險。微流控油包水液珠系統可以將微生物和抗生素封裝在微型生物反應器內，與周圍環境隔離，從而實現高通量的獨立檢測。本研究介紹了一種免標定介電泳（DEP）微流體平台的發展，用於基於電導度變化的DEP力（FDEP），在不需要光學分析的情況下，分選共封裝大腸桿菌（E. coli）和氨苄青黴素（Amp）的液珠與共封裝耐氨苄青黴素（AmpR）的大腸桿菌和Amp的液珠。稀釋的9.4%低電導率（LC）Luria–Bertani（LB）培養基加入170 mM甘露醇，可以維持大腸桿菌和AmpR大腸桿菌生長3小時，並允許Amp殺死幾乎所有的大腸桿菌，這顯著提高了LCLB電導率約100 μS/cm。因此，在施加的2 MHz和100 Vpp的電場下，產生正FDEP，透過兩液珠電導度差異，可以將未被殺死的AmpR大腸桿菌/9.4%LCLB/Amp和其中大多數被殺死的大腸桿菌/9.4%LCLB/Amp液珠進行分選。此外，當液珠中含有AmpR大腸桿菌的比例等於或高於50%時，分選比例顯著下降至約50%。基於電導率變化的DEP分選平台展示了通過分選率作為指標，探測未知細菌樣品中AmpR大腸桿菌比例的潛力。 |
| 英文摘要 | Antimicrobial resistance (AMR) has become a crucial global health issue. Antibiotic-resistant bacteria can survive after antibiotic treatments, lowering drug efficacy and increasing lethal risks. A microfluidic water-in-oil emulsion droplet system can entrap microorganisms and anti-biotics within the tiny bioreactor, separate from the surroundings, enabling independent assays that can be performed in a high-throughput manner. This study presents the development of a label-free dielectrophoresis (DEP)-based microfluidic platform to sort droplets that co-encapsulate Escherichia coli (E. coli) and ampicillin (Amp) and droplets that co-encapsulate Amp-resistant (AmpR) E. coli with Amp only based on the conductivity-dependent DEP force (FDEP) without the assistance of optical analyses. The 9.4% low conductivity (LC) Luria–Bertani (LB) broth diluted with 170 mM mannitol can maintain E. coli and AmpR E. coli growth for 3 h and allow Amp to kill almost all E. coli, which can significantly increase the LCLB conductivity by about 100 μS/cm. Therefore, the AmpR E. coli/9.4%LCLB/Amp where no cells are killed and the E. coli/9.4%LCLB/Amp-containing droplets where most of the cells are killed can be sorted based on this conductivity difference at an applied electric field of 2 MHz and 100 Vpp that generates positive FDEP. Moreover, the sorting ratio significantly decreased to about 50% when the population of AmpR E. coli was equal to or higher than 50% in droplets. The conductivity-dependent DEP-based sorting platform exhibits promising potential to probe the ratio of AmpR E. coli in an unknown bacterial sample by using the sorting ratio as an index. |
| 發表成果與AI計畫研究主題相關性 | 該研究已建立免標定式介電泳液珠分選平台，對含有氨苄青黴素液珠中大腸桿菌抗生素抗藥性進行分選測試。透過此平台可以延伸利用於禽畜養殖環境或是畜牧汙水處理場域中抗藥性細菌存在與否的評估，以監測抗藥性細菌在禽畜養殖場域或食品加工環境的污染威脅。目前已使用AI object detection技術針對每個液珠對電場極化的移動量進行分類，藉由物件檢測技術可辨識出液珠與其在為通道中的位置，從結果顯示可對不同抗藥性菌株含量的檢測更為靈敏。 |
