Polysaccharide-based liquid storage and transport media for non-refrigerated preservation of bacterial pathogens
Autor: | T. R. Pope, Kristin D. Victry, Brooke L. Deatherage Kaiser, Zachary C. Kennedy, Janine R. Hutchison, Kristin M. Omberg, Shelby M. Brooks, Kristie L. Oxford, Marvin G. Warner, Cynthia L. Warner |
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Jazyk: | angličtina |
Rok vydání: | 2019 |
Předmět: |
0301 basic medicine
Proteomics Staphylococcus Bacillus medicine.disease_cause Pathology and Laboratory Medicine Biochemistry Specimen Storage Nucleic Acids Medicine and Health Sciences Food science Staphylococcus Aureus Multidisciplinary Microbial Viability biology Chemistry Polysaccharides Bacterial Temperature Xanthomonas campestris Yersinia Bacillus anthracis Bacterial Pathogens Staphylococcus aureus Medical Microbiology Peptones Physical Sciences Medicine Pathogens medicine.drug Research Article Xanthomonas Yersinia Pestis Science 030106 microbiology Preservation Biological Bacillus Anthracis Research and Analysis Methods Microbiology 03 medical and health sciences Polysaccharides medicine Francisella novicida Cysteine Microbial Pathogens Xanthomonas Campestris Bacteria Organisms Chemical Compounds Biology and Life Sciences biology.organism_classification 030104 developmental biology Storage and Handling Nucleic acid Xanthan gum |
Zdroj: | PLoS ONE, Vol 14, Iss 9, p e0221831 (2019) PLoS ONE |
ISSN: | 1932-6203 |
Popis: | The preservation of biological samples for an extended time period of days to weeks after initial collection is important for the identification, screening, and characterization of bacterial pathogens. Traditionally, preservation relies on cold-chain infrastructure; however, in many situations this is impractical or not possible. Thus, our goal was to develop alternative bacterial sample preservation and transport media that are effective without refrigeration or external instrumentation. The viability, nucleic acid stability, and protein stability of Bacillus anthracis Sterne 34F2, Francisella novicida U112, Staphylococcus aureus ATCC 43300, and Yersinia pestis KIM D27 (pgm-) was assessed for up to 28 days. Xanthan gum (XG) prepared in PBS with L-cysteine maintained more viable F. novicida U112 cells at elevated temperature (40°C) compared to commercial reagents and buffers. Viability was maintained for all four bacteria in XG with 0.9 mM L-cysteine across a temperature range of 22-40°C. Interestingly, increasing the concentration to 9 mM L-cysteine resulted in the rapid death of S. aureus. This could be advantageous when collecting samples in the built environment where there is the potential for Staphylococcus collection and stabilization rather than other organisms of interest. F. novicida and S. aureus DNA were stable for up to 45 days upon storage at 22°C or 40°C, and direct analysis by real-time qPCR, without DNA extraction, was possible in the XG formulations. XG was not compatible with proteomic analysis via LC-MS/MS due to the high amount of residual Xanthomonas campestris proteins present in XG. Our results demonstrate that polysaccharide-based formulations, specifically XG with L-cysteine, maintain bacterial viability and nucleic acid integrity for an array of both Gram-negative and Gram-positive bacteria across ambient and elevated temperatures. |
Databáze: | OpenAIRE |
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