Continuous spore disruption using radially focused, high-frequency ultrasound

被引:50
作者
Chandler, DP [1 ]
Brown, J [1 ]
Bruckner-Lea, CJ [1 ]
Olson, L [1 ]
Posakony, GJ [1 ]
Stults, JR [1 ]
Valentine, NB [1 ]
Bond, LJ [1 ]
机构
[1] Pacific NW Natl Lab, Richland, WA 99352 USA
关键词
D O I
10.1021/ac010264j
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
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 1000-fold increase in intracellular DNA availability from ultrasonically disrupted spores, and liberated DNA was intact and available for subsequent detection.
引用
收藏
页码:3784 / 3789
页数:6
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