Microorganisms (May 2023)

DNA Capture and Enrichment: A Culture-Independent Approach for Characterizing the Genomic Diversity of Pathogenic <i>Leptospira</i> Species

  • Nathan E. Stone,
  • Ryelan F. McDonough,
  • Camila Hamond,
  • Karen LeCount,
  • Joseph D. Busch,
  • Katherine L. Dirsmith,
  • Sarai Rivera-Garcia,
  • Fred Soltero,
  • Laura M. Arnold,
  • Zachary Weiner,
  • Renee L. Galloway,
  • Linda K. Schlater,
  • Jarlath E. Nally,
  • Jason W. Sahl,
  • David M. Wagner

DOI
https://doi.org/10.3390/microorganisms11051282
Journal volume & issue
Vol. 11, no. 5
p. 1282

Abstract

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Because they are difficult to culture, obtaining genomic information from Leptospira spp. is challenging, hindering the overall understanding of leptospirosis. We designed and validated a culture-independent DNA capture and enrichment system for obtaining Leptospira genomic information from complex human and animal samples. It can be utilized with a variety of complex sample types and diverse species as it was designed using the pan-genome of all known pathogenic Leptospira spp. This system significantly increases the proportion of Leptospira DNA contained within DNA extracts obtained from complex samples, oftentimes reaching >95% even when some estimated starting proportions were Leptospira-positive human and animal samples. This, in turn, will lead to a better understanding of the overall genomic diversity and gene content of Leptospira spp. that cause leptospirosis, aiding epidemiology and the development of improved diagnostics and vaccines.

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