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评估磁性离子液体对非包膜病毒颗粒和游离病毒基因组RNA的分离及浓缩能力。

Evaluating the capacity of magnetic ionic liquids for separation and concentration of non-enveloped viral particles and free viral genomic RNA.

作者信息

Stoufer Sloane, Kim Minji, De Silva Shashini, Anderson Jared L, Brehm-Stecher Byron F, Moore Matthew D

机构信息

Department of Food Science, University of Massachusetts, Amherst, MA, USA.

Department of Chemistry, Iowa State University, Ames, IA, USA.

出版信息

Anal Bioanal Chem. 2025 Jan;417(2):435-445. doi: 10.1007/s00216-024-05662-6. Epub 2024 Nov 28.

Abstract

Magnetic ionic liquids (MILs) have proven effective as capture reagents for foodborne bacterial pathogens; however, there are currently no published studies regarding their use with foodborne, non-enveloped viruses. In this study, a protocol was evaluated for capture and recovery of bacteriophage MS2, a human norovirus surrogate, and purified viral genomic single stranded RNA (ssRNA) from an aqueous suspension using MILs. Transition metal-based MILs showed similar capture and recovery efficiency for both targets. A rare earth metal-based MIL showed much greater capture efficiency than the transition metal-based MILs, but displayed similar recovery. All tested MILs showed slightly higher capture and recovery efficiency for free RNA in comparison to intact virus, though overall trends were similar, and most MILs could recover both targets at as little as 10 PFU/mL intact MS2 or copies/mL purified RNA. A plaque assay confirmed that contact with MILs did not significantly reduce viral infectivity. Adjusting MIL volume gave no significant changes in capture or recovery, likely due to interplay between volume for the hydrophobic MIL and dispersion. Reducing the elution volume gave a slight increase in recovery, indicating MILs could be used for target enrichment after further optimization. MILs could also capture MS2 from romaine lettuce rinsate at comparable or even higher levels than from pure suspension, though loss in recovery was observed when the rinsate was prepared in an alkaline elution buffer. Overall, these results demonstrate the potential utility of MILs as concentration reagents for foodborne viruses, particularly for in-field applications.

摘要

磁性离子液体(MILs)已被证明是有效的食源性病原体捕获试剂;然而,目前尚无关于其用于食源性非包膜病毒的公开研究。在本研究中,评估了一种使用MILs从水悬浮液中捕获和回收噬菌体MS2(一种人类诺如病毒替代物)以及纯化病毒基因组单链RNA(ssRNA)的方案。基于过渡金属的MILs对两个目标显示出相似的捕获和回收效率。一种基于稀土金属的MIL显示出比基于过渡金属的MILs更高的捕获效率,但回收效率相似。与完整病毒相比,所有测试的MILs对游离RNA的捕获和回收效率略高,尽管总体趋势相似,并且大多数MILs在低至10 PFU/mL完整MS2或拷贝/mL纯化RNA时都能回收两个目标。噬菌斑测定证实与MILs接触不会显著降低病毒感染性。调整MIL体积对捕获或回收没有显著影响,这可能是由于疏水性MIL的体积与分散之间的相互作用。减少洗脱体积会使回收率略有提高,这表明在进一步优化后MILs可用于目标富集。MILs还可以从生菜冲洗液中捕获MS2,其水平与从纯悬浮液中捕获的相当甚至更高,尽管当在碱性洗脱缓冲液中制备冲洗液时观察到回收率有所损失。总体而言,这些结果证明了MILs作为食源性病毒浓缩试剂的潜在用途,特别是在现场应用中。

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