Document Detail


Mechanical disruption of lysis-resistant bacterial cells by use of a miniature, low-power, disposable device.
MedLine Citation:
PMID:  21543569     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Molecular detection of microorganisms requires microbial cell disruption to release nucleic acids. Sensitive detection of thick-walled microorganisms such as Bacillus spores and Mycobacterium cells typically necessitates mechanical disruption through bead beating or sonication, using benchtop instruments that require line power. Miniaturized, low-power, battery-operated devices are needed to facilitate mechanical pathogen disruption for nucleic acid testing at the point of care and in field settings. We assessed the lysis efficiency of a very small disposable bead blender called OmniLyse relative to the industry standard benchtop Biospec Mini-BeadBeater. The OmniLyse weighs approximately 3 g, at a size of approximately 1.1 cm(3) without the battery pack. Both instruments were used to mechanically lyse Bacillus subtilis spores and Mycobacterium bovis BCG cells. The relative lysis efficiency was assessed through real-time PCR. Cycle threshold (C(T)) values obtained at all microbial cell concentrations were similar between the two devices, indicating that the lysis efficiencies of the OmniLyse and the BioSpec Mini-BeadBeater were comparable. As an internal control, genomic DNA from a different organism was spiked at a constant concentration into each sample upstream of lysis. The C(T) values for PCR amplification of lysed samples using primers specific to this internal control were comparable between the two devices, indicating negligible PCR inhibition or other secondary effects. Overall, the OmniLyse device was found to effectively lyse tough-walled organisms in a very small, disposable, battery-operated format, which is expected to facilitate sensitive point-of-care nucleic acid testing.
Authors:
Peter E Vandeventer; Kris M Weigel; Jose Salazar; Barbara Erwin; Bruce Irvine; Robert Doebler; Ali Nadim; Gerard A Cangelosi; Angelika Niemz
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2011-05-04
Journal Detail:
Title:  Journal of clinical microbiology     Volume:  49     ISSN:  1098-660X     ISO Abbreviation:  J. Clin. Microbiol.     Publication Date:  2011 Jul 
Date Detail:
Created Date:  2011-07-01     Completed Date:  2011-09-26     Revised Date:  2013-06-30    
Medline Journal Info:
Nlm Unique ID:  7505564     Medline TA:  J Clin Microbiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2533-9     Citation Subset:  IM    
Affiliation:
Keck Graduate Institute of Applied Life Sciences, 535 Watson Drive, Claremont, California 91711, USA.
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MeSH Terms
Descriptor/Qualifier:
Bacillus subtilis / genetics*
Bacteriological Techniques / methods*
Bacteriolysis*
DNA, Bacterial / isolation & purification*
Humans
Mycobacterium bovis / genetics*
Grant Support
ID/Acronym/Agency:
AI090831/AI/NIAID NIH HHS; R01 AI090831/AI/NIAID NIH HHS; R01 AI090831-02/AI/NIAID NIH HHS; R01 AI090831-03/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/DNA, Bacterial
Comments/Corrections

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