Rapid sample preparation with Lyse-It® for Listeria monocytogenes and Vibrio cholerae
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Date
2018-07-25
Type of Work
Department
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Citation of Original Publication
Santaus TM, Li S, Ladd P, Harvey A, Cole S, et al. (2018) Rapid sample preparation with Lyse-It® for Listeria monocytogenes and Vibrio cholerae. PLOS ONE 13(7): e0201070. https://doi.org/10.1371/journal.pone.0201070
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Attribution 4.0 International (CC BY 4.0)
Attribution 4.0 International (CC BY 4.0)
Subjects
Abstract
Sample preparation is a leading bottleneck in rapid detection of pathogenic bacteria. Here, we use Lyse-It® for bacterial cellular lysis, genomic DNA fragmentation, and protein release and degradation for both Listeria monocytogenes and Vibrio cholerae. The concept of Lyse-It® employs a conventional microwave and Lyse-It® slides for intensely focused microwave irradiation onto the sample. High microwave power and a <60 second irradiation time allow for rapid cellular lysis and subsequent intracellular component release. The pathogenic bacteria are identified by quantitative polymerase chain reaction (qPCR), which subsequently demonstrates the viability of DNA for amplification post microwave-induced lysis. Intracellular component release, degradation, and detection of L. monocytogenes and V. cholerae has been performed and shown in this paper. These results demonstrate a rapid, low-cost, and efficient way for bacterial sample preparation on both food and water-borne Gram-positive and -negative organisms alike.