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Synthesis and application of superabsorbent polymer microspheres for rapid concentration and quantification of microbial pathogens in ambient water.
Separation and Purification Technology ( IF 8.1 ) Pub Date : 2020-05-15 , DOI: 10.1016/j.seppur.2020.116540 Xunyi Wu 1 , Xiao Huang 1 , Yanzhe Zhu 1 , Jing Li 1 , Michael R Hoffmann 1
Separation and Purification Technology ( IF 8.1 ) Pub Date : 2020-05-15 , DOI: 10.1016/j.seppur.2020.116540 Xunyi Wu 1 , Xiao Huang 1 , Yanzhe Zhu 1 , Jing Li 1 , Michael R Hoffmann 1
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Even though numerous methods have been developed for the detection and quantification of waterborne pathogens, the application of these methods is often hindered by the very low pathogen concentrations in natural waters. Therefore, rapid and efficient sample concentration methods are urgently needed. Here we present a novel method to pre-concentrate microbial pathogens in water using a portable 3D-printed system with super-absorbent polymer (SAP) microspheres, which can effectively reduce the actual volume of water in a collected sample. The SAP microspheres absorb water while excluding bacteria and viruses by size exclusion and charge repulsion. To improve the water absorption capacity of SAP in varying ionic strength waters (0-100 mM), we optimized the formulation of SAP to 180 g⋅L-1 Acrylamide, 75 g⋅L-1 Itaconic Acid and 4.0 g⋅L-1 Bis-Acrylamide for the highest ionic strength water as a function of the extent of cross-linking and the concentration of counter ions. Fluorescence microscopy and double-layer agar plating respectively showed that the 3D-printed system with optimally-designed SAP microspheres could rapidly achieve a 10-fold increase in the concentration of Escherichia coli (E. coli) and bacteriophage MS2 within 20 min with concentration efficiencies of 87% and 96%, respectively. Fold changes between concentrated and original samples from qPCR and RT-qPCR results were found to be respectively 11.34-22.27 for E. coli with original concentrations from 104 to 106 cell·mL-1, and 8.20-13.81 for MS2 with original concentrations from 104 to 106 PFU·mL-1. Furthermore, SAP microspheres can be reused for 20 times without performance loss, significantly decreasing the cost of our concentration system.
中文翻译：
超吸收性聚合物微球的合成和应用，用于快速浓缩和定量环境水中的微生物病原体。
尽管已开发出许多方法来检测和定量水生病原体，但由于天然水中的病原体浓度非常低，这些方法的应用常常受到阻碍。因此，迫切需要快速有效的样品浓缩方法。在这里，我们介绍了一种使用便携式3D打印系统和高吸收性聚合物（SAP）微球预先浓缩水中微生物病原体的新方法，该方法可以有效地减少收集样品中的实际水量。SAP微球吸收水，同时通过尺寸排阻和排斥电荷排除细菌和病毒。为了提高在不同离子强度的水中（0-100 mM）中SAP的吸水能力，我们将SAP的配方优化为180g⋅L-1丙烯酰胺，75g⋅L-1衣康酸和4。0g⋅L-1双丙烯酰胺，水的最高离子强度取决于交联程度和抗衡离子浓度。荧光显微镜和双层琼脂平板分别显示，具有优化设计的SAP微球的3D打印系统可以在20分钟内迅速实现大肠杆菌（E. coli）和噬菌体MS2的浓度增加10倍，并且具有较高的浓度效率分别为87％和96％。qPCR和RT-qPCR结果的浓缩样品和原始样品之间的倍数变化分别为：原始浓度为104-106细胞·mL-1的大肠杆菌为11.34-22.27，原始浓度为104的MS2为8.20-13.81。至106 PFU·mL-1。此外，SAP微球可以重复使用20次而不会造成性能损失，
更新日期：2020-01-13
中文翻译：
超吸收性聚合物微球的合成和应用，用于快速浓缩和定量环境水中的微生物病原体。
尽管已开发出许多方法来检测和定量水生病原体，但由于天然水中的病原体浓度非常低，这些方法的应用常常受到阻碍。因此，迫切需要快速有效的样品浓缩方法。在这里，我们介绍了一种使用便携式3D打印系统和高吸收性聚合物（SAP）微球预先浓缩水中微生物病原体的新方法，该方法可以有效地减少收集样品中的实际水量。SAP微球吸收水，同时通过尺寸排阻和排斥电荷排除细菌和病毒。为了提高在不同离子强度的水中（0-100 mM）中SAP的吸水能力，我们将SAP的配方优化为180g⋅L-1丙烯酰胺，75g⋅L-1衣康酸和4。0g⋅L-1双丙烯酰胺，水的最高离子强度取决于交联程度和抗衡离子浓度。荧光显微镜和双层琼脂平板分别显示，具有优化设计的SAP微球的3D打印系统可以在20分钟内迅速实现大肠杆菌（E. coli）和噬菌体MS2的浓度增加10倍，并且具有较高的浓度效率分别为87％和96％。qPCR和RT-qPCR结果的浓缩样品和原始样品之间的倍数变化分别为：原始浓度为104-106细胞·mL-1的大肠杆菌为11.34-22.27，原始浓度为104的MS2为8.20-13.81。至106 PFU·mL-1。此外，SAP微球可以重复使用20次而不会造成性能损失，
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