engleski

A novel application of CIM monolithic supports for concentrating Potato spindle tuber viroid from water samples

Potato spindle tuber viroid (PSTVd) is the causal agent of a number of agriculturally important diseases. It is a single-stranded, circular and uncapsidated RNA molecule with 359 nucleotides and no coding capacity. Because of its complex secondary/tertiary structure it is very stable ex vivo and it is easily transmitted mechanically by contaminated hands, tools, machinery, etc. In this work, we describe the development and optimization of a method for concentrating PSTVd using CIM monolithic supports. The ion-exchange chromatography on CIMacTM DEAE columns resulted in up to 30% PSTVd recovery whilst the hydrophobic interaction chromatography on CIMacTM C4 columns improved it up to 60%. Based on these results, a CIM® C4 HLD-1 (High Ligand Density) tube monolithic column was selected for further experiments. One-liter-water samples were mixed with different viroid quantities and loaded onto the column. By using RT-qPCR, the viroid RNA was quantified in the elution fraction (≈ 5 mL) indicating that 70% of the viroid was recovered and concentrated by at least two orders of magnitude. This approach would be helpful in screening irrigation waters and/or hydroponic systems’ nutrient solutions for the presence of PSTVd.

viroid; chromatography; monolithic support; concentrating

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