Continuous-Flow, Rapid Lysis Devices for Biodefense Nucleic Acid Diagnostic Systems

Abstract

Two mechanical lysis devices have been developed as compact, robust components to provide rapid sample preparation for nucleic acid diagnostic systems. One such component, known as the Micro Bead-Beater™ (μBB™, BBTM, Claremont BioSolutions, Upland, CA), is a compact device that is capable of ultrarapid lysis (>90% lysis in 30 s) of micro volumes (<80 μL) of Bacillus spores in a continuous-flow format or in a disposable single-tube format. The μBB is also capable of processing much larger volumes of solutions containing spores or vegetative cells using a continuous-flow mode. A second mechanical lysis device designed as a disposable component is the microfluidic bead blender, which uses a small electric motor to spin vanes within the bead-laden solution. DNA quantification results using dsDNA-binding fluorescence dyes and real-time PCR are presented, comparing the lysis of Bacillus subtilis spores using the μBB™ with other well-known lysis techniques. Nanoscale imaging results obtained using scanning electron microscopy and transmission electron microscopy on B. subtilis spores lyzed using the μBB™ are also presented