title: Deglycosylation of Tropheryma whipplei biofilm and discrepancies between diagnostic results during Whipple's disease progression.
abstract: Whipple's disease is a systemic infectious disease associated with the bacterium Tropheryma whipplei. Numerous reports have presented puzzling discrepancies between diagnosis methods. We addressed this confusion using fluorescent in situ hybridization and immunofluorescence assays to evaluate 34 duodenal biopsies and 1 lymph node biopsy from Whipple's patients. We showed the presence of bacteria in both CK20(+) epithelial cells and CD68(+) macrophages. Bacteria are found embedded in a biofilm hindering the detection of T. whipplei. Only after treatment of biopsies by glycosidases, co-localization of T. whipplei RNA/DNA with bacterial proteins was restored. Moreover, using 13 bronchoalveolar lavages and 7 duodenal biopsies, we found that hydrolysis of the biofilm weakened the bacteria, facilitated bacterial DNA extraction and improved the sensitivity of qPCR detection by up to 1000x opening new perspectives for diagnostic and scientific approaches.
authors.affiliation: Aix Marseille Université, URMITE, UM63, CNRS 7278, IRD 198, Inserm 1095, 13005 Marseille, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Audoly
authors.affiliation: Aix Marseille Université, URMITE, UM63, CNRS 7278, IRD 198, Inserm 1095, 13005 Marseille, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Fenollar
authors.affiliation: Aix Marseille Université, URMITE, UM63, CNRS 7278, IRD 198, Inserm 1095, 13005 Marseille, France.
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Lagier
authors.affiliation: Aix Marseille Université, URMITE, UM63, CNRS 7278, IRD 198, Inserm 1095, 13005 Marseille, France.
authors.forename: Hubert
authors.initials: H
authors.lastname: Lepidi
authors.affiliation: Aix Marseille Université, URMITE, UM63, CNRS 7278, IRD 198, Inserm 1095, 13005 Marseille, France.
authors.forename: Didier
authors.initials: D
authors.lastname: Raoult
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology / Q000627:therapeutic use; D001433:Bacteriolysis; D018441:Biofilms; D001706:Biopsy; D018450:Disease Progression; D004386:Duodenum / Q000382:microbiology / Q000473:pathology; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000494:pharmacology; D006031:Glycosylation; D006801:Humans; D007413:Intestinal Mucosa / Q000382:microbiology / Q000473:pathology; D008198:Lymph Nodes / Q000382:microbiology / Q000473:pathology; D018613:Microscopy, Confocal; D054851:Tropheryma / Q000502:physiology*; D008061:Whipple Disease / Q000175:diagnosis* / Q000188:drug therapy / Q000382:microbiology
chemical_list: D000900:Anti-Bacterial Agents; D006026:Glycoside Hydrolases