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
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
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje