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Multi-step optimization of the filtration method for the isolation of Campylobacter species from stool samples

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Abstract

The filtration method (FM) is the most effective isolation technique for Epsilobacteriaceae from stool samples. FM’s different adaptations make it difficult to compare data between studies. This study was performed in three phases to optimize FM from a routine laboratory perspective. In July–September 2014 (part I), FM was performed on Mueller–Hinton agar containing 5% sheep blood and Columbia agar containing 5% sheep blood. In July 2016 (part II), FM was performed using 0.60-μm pore size polycarbonate filters (0.6-PC filter) and 0.45-μm pore size cellulose acetate filters (0.45-AC filter); in January 2018 (part III), the addition of hydrogen to incubators was studied. On 1146 stools analyzed in part I, the positive samples that showed no growth on the Butzler medium (n = 22/72, 30.6%) had improved growth of Epsilobacteriaceae when using the Columbia instead of the Mueller–Hinton medium (21/22 strains vs. 11/22, p < 0.05). In part II, on 718 stools, 91 strains grew with FM (12.7%), more with 0.6-PC filter (90/91) than with 0.45-AC filter (44/91) (p < 0.05). In part III, 578 stools were cultured, 98 Epsilobacteriaceae strains grew with FM, and 7% hydrogen finding significantly more Epsilobacteriaceae than without hydrogen (90/98, 91.8%, vs. 72/98, 73.5%; p < 0.05). The use of a Columbia medium containing 5% sheep blood with 0.6-PC filters incubated at 37 °C in a 7% hydrogen-enriched atmosphere led to an almost fourfold increase in the isolation rate of Epsilobacteriaceae among the studied combinations. Reference centers for Campylobacter should use standardized protocols to enable the comparison of prevalence in space and time.

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Acknowledgments

This work was supported by The Belgian Kids’ Fund for Pediatric Research.

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Author notes

  1. Marie Hallin and Delphine Martiny act as equivalent co-senior authors

Authors and Affiliations

  1. Division of Infection Prevention and Control, Hôpital Universitaire des Enfants Reine Fabiola, Brussels, Belgium

    Anne Tilmanne

  2. Division of Pediatric Infectious Diseases, CHU Sainte Justine, Montreal, Quebec, Canada

    Anne Tilmanne & Caroline Quach

  3. Department of Microbiology, Laboratoire Hospitalier Universitaire de Bruxelles – Universitair Laboratorium Brussel (LHUB-ULB), Université Libre de Bruxelles (ULB), Brussels, Belgium

    Helga Marisca Kandet Yattara, Margaux Herpol, Linda Vlaes, Patricia Retore, Marie Hallin & Delphine Martiny

  4. National Reference Centre for Campylobacter, CHU Saint-Pierre, Brussels, Belgium

    Margaux Herpol, Linda Vlaes, Patricia Retore, Olivier Vandenberg & Delphine Martiny

  5. Department of Microbiology, Infectious Diseases & Immunology, Université de Montréal, Montreal, Canada

    Caroline Quach

  6. Innovation and Business Development Unit, LHUB-ULB, Pole Hospitalier Universitaire de Bruxelles, Université Libre de Bruxelles, Brussels, Belgium

    Olivier Vandenberg

  7. Centre for Environmental Health and Occupational Health, School of Public Health, Université Libre de Bruxelles (ULB), Brussels, Belgium

    Olivier Vandenberg

  8. Faculté de Médecine et Pharmacie, Université de Mons (UMONS), Mons, Belgium

    Delphine Martiny

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  1. Anne Tilmanne
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Correspondence to Anne Tilmanne.

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Tilmanne, A., Kandet Yattara, H.M., Herpol, M. et al. Multi-step optimization of the filtration method for the isolation of Campylobacter species from stool samples. Eur J Clin Microbiol Infect Dis 38, 859–864 (2019). https://doi.org/10.1007/s10096-019-03479-1

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  • DOI: https://doi.org/10.1007/s10096-019-03479-1

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