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Applied Microbiology and Biotechnology

, Volume 98, Issue 5, pp 2243–2254 | Cite as

Optical forward-scattering for identification of bacteria within microcolonies

  • Pierre R. Marcoux
  • Mathieu Dupoy
  • Antoine Cuer
  • Joe-Loïc Kodja
  • Arthur Lefebvre
  • Florian Licari
  • Robin Louvet
  • Anil Narassiguin
  • Frédéric Mallard
Methods and protocols

Abstract

The development of methods for the rapid identification of pathogenic bacteria is a major step towards accelerated clinical diagnosis of infectious diseases and efficient food and water safety control. Methods for identification of bacterial colonies on gelified nutrient broth have the potential to bring an attractive solution, combining simple optical instrumentation, no need for sample preparation or labelling, in a non-destructive process. Here, we studied the possibility of discriminating different bacterial species at a very early stage of growth (6 h of incubation at 37 °C), on thin layers of agar media (1 mm of Tryptic Soy Agar), using light forward-scattering and learning algorithms (Bayes Network, Continuous Naive Bayes, Sequential Minimal Optimisation). A first database of more than 1,000 scatterograms acquired on 7 gram-negative strains yielded a recognition rate of nearly 80 %, after only 6 h of incubation. We investigated also the prospect of identifying different strains from a same species through forward scattering. We discriminated, thus, four strains of Escherichia coli with a recognition rate reaching 82 %. Finally, we show the discrimination of two species of coagulase-negative Staphylococci (S. haemolyticus and S. cohnii), on a commercial selective pre-poured medium used in clinical diagnosis (ChromID MRSA, bioMérieux), without opening lids during the scatterogram acquisition. This shows the potential of this method—non-invasive, preventing cross-contaminations and requiring minimal dish handling—to provide early clinically-relevant information in the context of fully automated microbiology labs.

Keywords

Bacteria Identification Forward-scattering Label-free Microcolonies Non-invasive technique 

Notes

Acknowledgments

We are grateful to Frédéric Pinston, Quentin Jossso and Sylvain Orenga from bioMérieux for helpful discussions. Charles-Edmond Bichot (Ecole Centrale de Lyon) is gratefully acknowledged for assistance in java programming.

Supplementary material

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Pierre R. Marcoux
    • 1
  • Mathieu Dupoy
    • 1
  • Antoine Cuer
    • 2
  • Joe-Loïc Kodja
    • 2
  • Arthur Lefebvre
    • 2
  • Florian Licari
    • 2
  • Robin Louvet
    • 2
  • Anil Narassiguin
    • 2
  • Frédéric Mallard
    • 3
  1. 1.Department of Technology for Biology and HealthcareCEA-LETI MINATECGrenobleFrance
  2. 2.Ecole Centrale de LyonEcullyFrance
  3. 3.bioMérieux SA, Innovation & Systems/Technology Research/Sample Prep & Processing LabGrenobleFrance

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