Continuous spore disruption using radially focused, high-frequency ultrasound.
We report on the development of a novel, continuous-flow, radially focused ultrasonic disruptor capable of lysing Bacillus spores in the absence of added chemical denaturants, enzymes, or microparticles. Greater than 99% disruption was achieved for Bacillus globigii spores and Escherichia coli and Bacillus subtilis vegetative cells with sample residence times of 62, 12, and 12 s, respectively. Microscopic and SEM images indicated that at equivalent power levels, the incidence of cell death or loss of viability typically exceeded the efficiency of (visible) cell lysis. However, semiquantitative PCR showed up to a 1,000-fold increase in intracellular DNA availability from ultrasonically disrupted spores, and liberated DNA was intact and available for subsequent detection.
Chandler
D P
DP
Pacific Northwest National Laboratory, Richland, Washington 99352, USA. dp.chandler@pnl.gov
Brown
J
J
Bruckner-Lea
C J
CJ
Olson
L
L
Posakony
G J
GJ
Stults
J R
JR
Valentine
N B
NB
Bond
L J
LJ
D001407:Bacillus; D002470:Cell Survival / Q000528:radiation effects; D004247:DNA / Q000737:chemistry; D004926:Escherichia coli / Q000166:cytology; D008855:Microscopy, Electron, Scanning; D016133:Polymerase Chain Reaction; D013171:Spores, Bacterial / Q000166:cytology* / Q000528:radiation effects* / Q000648:ultrastructure; D014465:Ultrasonics
D004247:DNA