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

Journal Article · Thu Oct 01 00:00:00 EDT 2009 · JALA. Journal of the Association for Laboratory Automation, 14(5):277-284

DOI:https://doi.org/10.1016/j.jala.2009.04.007· OSTI ID:979509

Warner, Cynthia L; Bruckner-Lea, Cindy J; Grate, Jay W; Straub, Tim M; Posakony, Gerald J; Valentine, Nancy B; Ozanich, Richard M; Bond, Leonard J; Matzke, Melissa M; Dockendorff, Brian P; Valdez, Catherine O; Valdez, Patrick LJ; Owsley, Stanley L

An automated, flow-through spore lysis instrument that is capable of rapidly disrupting bacterial spores is described. The system utilizes a flow-through chamber that allows for direct injection of the sample without the need for a chemical or enzymatic pre-treatment step to soften the spore coat prior to 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, which showed at least a 25-fold increase in amplifiable DNA following ultrasonic treatment, and dilution-to-extinction PCR, which suggests up to a 100-1000-fold increase. The modular design of the ultrasonic system and integrated fluidics allow it to be incorporated into multi-step sample treatment and detection systems.

• Research Organization: Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 979509
• Report Number(s): PNNL-SA-58098; 30400; TRN: US201010%%732
• Journal Information: JALA. Journal of the Association for Laboratory Automation, 14(5):277-284, Vol. 14, Issue 5
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE
BACILLUS
BACILLUS SUBTILIS
BACTERIAL SPORES
DESIGN
DETECTION
DNA
LYSIS
SPORES
TRANSDUCERS
ULTRASONIC WAVES
spore
Bacillus
ultrasound
sonication
sequential injection analysis
automation
Environmental Molecular Sciences Laboratory