A Flow-Through Ultrasonic Lysis Module for the Disruption of Bacterial Spores

被引:11
|
作者
Warner, Cynthia L. [1 ]
Bruckner-Lea, Cynthia J. [1 ]
Grate, Jay W. [1 ]
Straub, Timothy [1 ]
Posakony, Gerald J. [1 ]
Valentine, Nancy [1 ]
Ozanich, Richard [1 ]
Bond, Leonard J. [1 ]
Matzke, Melissa M. [1 ]
Dockendorff, Brian [1 ]
Valdez, Catherine [1 ]
Valdez, Patrick [1 ]
Owsley, Stanley [1 ]
机构
[1] Pacific NW Natl Lab, Richland, WA 99352 USA
来源
JALA | 2009年 / 14卷 / 05期
关键词
spore; Bacillus; ultrasound; sonication; sequential injection analysis; automation; SAMPLE PREPARATION METHOD; RENEWABLE MICROCOLUMNS; DNA ANALYSIS; CELL-LYSIS; SONICATION; INJECTION; TIME; PCR; AMPLIFICATION; PURIFICATION;
D O I
10.1016/j.jala.2009.04.007
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
An automated, flow-through ultrasonic lysis module that is capable of disrupting bacterial spores to increase the DNA available for biodetection is described. The system uses a flow-through chamber that allows for direct injection of the sample without the need for a chemical or enzymatic pretreatment step to disrupt the spore coat before lysis. Lysis of Bacillus subtilis spores, a benign simulant of Bacillus anthracis, is achieved by flowing the sample through a tube whose axis is parallel to the faces of two transducers that deliver 10 W cm(-2) to the surface of the tube at 1.4-MHz frequency. Increases in amplifiable DNA were assessed by real-time PCR analysis that showed at least a 25-fold increase in amplifiable DNA after ultrasonic treatment with glass beads, compared with controls with no ultrasonic power applied. The ultrasonic system and integrated fluidics are designed as a module that could be incorporated into multistep, automated sample treatment and detection systems for pathogens. (JALA 2009;14:277-84)
引用
收藏
页码:277 / 284
页数:8
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