Novel sampling technique maintaining the two-dimensional organization of microbes during cultivation from chronic wounds: The Imprint method

Anne Kristine Servais Iversen, Blaine Gabriel Fritz, Mads Joachim Hansen, Klaus Kirketerp-Møller, Tim Holm Jakobsen, Thomas Bjarnsholt, Mads Lichtenberg*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

This study aimed to develop and validate “the Imprint method,”, a technique for sampling microbes from chronic wounds while preserving their two-dimensional spatial organization. We used nylon filters to sample bacteria and compared with sampling using Eswabs in 12 patients. The Imprint method identified a mean of 0.93 unique species more than Eswab (4.3 ± 2.2 and 3.4 ± 1.4 unique species, respectively; mean ± SD; n = 30). Accuracy between the Eswab and the Imprint method was 93.2% and in cases of disagreement between methods, Imprint had a higher sensitivity in 6/8 of the most prevalent species. In vitro validation confirmed that the Imprint method could transfer bacterial colonies while replicating their two-dimensional organization and the area covered by bacteria on the plate sampled. Clinical testing demonstrated that the imprint method is a rapid and feasible technique that identified more unique bacterial species than Eswab with a good agreement between methods but that Imprint was better at detecting important pathogens such as Staphylococcus aureus and Pseudomonas aeruginosa. The Imprint method is a novel technique that cultures and records the two-dimensional organization of microbes, providing an alternative or supplement to conventional surface culture using Eswab.

Original languageEnglish
JournalAPMIS
Volume132
Pages (from-to)210-220
ISSN0903-4641
DOIs
Publication statusPublished - 2024

Bibliographical note

Publisher Copyright:
© 2024 The Authors. APMIS published by John Wiley & Sons Ltd on behalf of Scandinavian Societies for Pathology, Medical Microbiology and Immunology.

Keywords

  • bacteria
  • chronic infection
  • Chronic wounds
  • culture
  • Imprint
  • techniques

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