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PLoS One. 2019 Jul 23;14(7):e0220102. doi: 10.1371/journal.pone.0220102. eCollection 2019.

A comparison of Lyse-It to other cellular sample preparation, bacterial lysing, and DNA fragmentation technologies.

Author information

1
Chemistry and Biochemistry Department, University of Maryland, Baltimore County, Baltimore, MD, United States of America.
2
Institute of Fluorescence, University of Maryland, Baltimore County, Baltimore, MD, United States of America.
3
Epidemiology and Public Health Department, University of Maryland School of Medicine, Baltimore, MD, United States of America.
4
Center for Vaccine Development and Global Health, University of Maryland School of Medicine, Baltimore, MD, United States of America.

Abstract

The ability for safe and rapid pathogenic sample transportation and subsequent detection is an increasing challenge throughout the world. Herein, we describe and use bead-beating, vortex, sonication, 903 protein saver cards, and Lyse-It methods, aiming to inactivate Gram-positive and -negative bacteria with subsequent genome DNA (quantitative Polymerase Chain Reaction) qPCR detection. The basic concepts behind the four chosen technologies is their versatility, cost, and ease of use in developed and underdeveloped countries. The four methods target the testing of bacterial resilience, cellular extraction from general and complex media and subsequent DNA extraction for qPCR detection and amplification. These results demonstrate that conventional high temperature heating, 903 protein saver cards, and Lyse-It are all viable options for inactivating bacterial growth for safe shipping. Additionally, Lyse-It was found to be particularly useful as this technology can inactivate bacteria, extract cells from 903 protein saver cards, lyse bacterial cells, and additionally keep genomic DNA viable for qPCR detection.

Conflict of interest statement

This work is supported by the UMBC-UMB Cholera joint training grant and the National Institutes of Health UMBC Graduate Training Chemistry Biology Interface Fellowship (T32GM066706-15). We note that the University of Maryland, Baltimore County (UMBC) has filed and has had issued several patents related to microwave-based lysing and DNA fragmentation. Those patents are licensed to Lyse-It LLC, a Maryland-based biotechnology company to which Dr. Geddes currently owns stock. Patent #: US US9500590B2. “Assays for pathogen detection using microwaves for lysing and accelerating metal-enhanced fluorescence”, Chris D. Geddes. Patent #: US10294451B2 “Flow and static lysing systems and methods for ultra-rapid isolation and fragmentation of biological materials by microwave irradiation”, Chris D. Geddes. Lyse-It LLC was not involved in the study design of the material within this paper,nor do any of the author receive any salaries or consultancies from Lyse-It LLC.

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