Rapid Extraction of Viral Nucleic Acids Using Rotating Blade Lysis and Magnetic Beads

Abstract

The complex and lengthy protocol of current viral nucleic acid extraction processes limits their use outside laboratory settings. Here, we describe a rapid and reliable method for extracting nucleic acids from viral samples using a rotating blade and magnetic beads. The viral membrane can be instantly lysed using a high-speed rotating blade, and nucleic acids can be immediately isolated using a silica magnetic surface. The process was completed within 60 s by this method. Routine washing and eluting processes were subsequently conducted within 5 min. The results achieved by this method were comparable to those of a commercially available method. When the blade-based lysis and magnetic bead adsorption processes were performed separately, the RNA recovery rate was very low, and the Ct value was delayed compared to simultaneous lysis and RNA adsorption. Overall, this method not only dramatically shortens the conventional extraction time but also allows for its convenient use outside the laboratory, such as at remote field sites and for point-of-care testing.

Development of a Magnetic Bead-Based Method for Specific Detection of Enterococcus faecalis Using C-Terminal Domain of ECP3 Phage Endolysin

“…The advantage of Ni-NTA MBs is their convenient coating procedure, high sedimentation ratio, and low surface/volume ratio; however, their unstable CBD binding precluded their further use for the isolation of whole bacterial cells at low concentrations (unpublished data). In contrast, eMBs are small, hydrophilic, polystyrene-coated particles whose specific surface chemistry allows for covalent protein immobilization [ 24 ]. These particles offer further advantages, including a low sedimentation ratio (increased availability of the coated surface), high stability, and high surface/volume ratio [ 25 ].…”