title: Fluorescence in situ hybridization for the identification of Treponema pallidum in tissue sections.
abstract: Syphilis is often called the great imitator because of its frequent atypical clinical manifestations that make the disease difficult to recognize. Because Treponema pallidum subsp. pallidum, the infectious agent of syphilis, is yet uncultivated in vitro, diagnosis is usually made using serology; however, in cases where serology is inconclusive or in patients with immunosuppression where these tests may be difficult to interpret, the availability of a molecular tool for direct diagnosis may be of pivotal importance. Here we present a fluorescence in situ hybridization (FISH) assay that simultaneously identifies and analyzes spatial distribution of T. pallidum in histological tissue sections. For this assay the species-specific FISH probe TPALL targeting the 16S rRNA of T. pallidum was designed in silico and evaluated using T. pallidum infected rabbit testicular tissue and a panel of non-syphilis spirochetes as positive and negative controls, respectively, before application to samples from four syphilis-patients. In a HIV positive patient, FISH showed the presence of T. pallidum in inguinal lymph node tissue. In a patient not suspected to suffer from syphilis but underwent surgery for phimosis, numerous T. pallidum cells were found in preputial tissue. In two cases with oral involvement, FISH was able to differentiate T. pallidum from oral treponemes and showed infection of the oral mucosa and tonsils, respectively. The TPALL FISH probe is now readily available for in situ identification of T. pallidum in selected clinical samples as well as T. pallidum research applications and animal models.
authors.affiliation: Institute for Microbiology and Hygiene, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Annett
authors.initials: A
authors.lastname: Petrich
authors.affiliation: Institute for Microbiology and Hygiene, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Pablo
authors.initials: P
authors.lastname: Rojas
authors.affiliation: Biofilmzentrum, Deutsches Herzzentrum Berlin, Consultant Laboratory for Tropheryma whipplei, Berlin, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Schulze
authors.affiliation: Institute for Pathology, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Loddenkemper
authors.affiliation: Department of Medicine, Division of Allergy and Infectious Diseases, and Department of Global Health, University of Washington, Seattle, WA 98104, USA.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Giacani
authors.affiliation: Department of Gastroenterology, Infectiology and Rheumatology, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Schneider
authors.affiliation: Department of Oral Medicine, Dental Radiology and Oral Surgery, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Moritz
authors.initials: M
authors.lastname: Hertel
authors.affiliation: Institute for Microbiology and Hygiene, Charité-Universitätsmedizin Berlin, Berlin, Germany; Biofilmzentrum, Deutsches Herzzentrum Berlin, Consultant Laboratory for Tropheryma whipplei, Berlin, Germany.
authors.forename: Judith
authors.initials: J
authors.lastname: Kikhney
authors.affiliation: Biofilmzentrum, Deutsches Herzzentrum Berlin, Consultant Laboratory for Tropheryma whipplei, Berlin, Germany. Electronic address: moter@dhzb.de.
authors.forename: Annette
authors.initials: A
authors.lastname: Moter
mesh_terms: D000328:Adult; D000368:Aged; D000818:Animals; D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D004195:Disease Models, Animal; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008198:Lymph Nodes / Q000382:microbiology / Q000473:pathology; D008297:Male; D008875:Middle Aged; D009061:Mouth Mucosa / Q000382:microbiology / Q000473:pathology; D015345:Oligonucleotide Probes / Q000235:genetics; D014066:Palatine Tonsil / Q000382:microbiology / Q000473:pathology; D057089:Pathology, Molecular / Q000379:methods*; D010413:Penis / Q000382:microbiology / Q000473:pathology; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D011817:Rabbits; D013587:Syphilis / Q000175:diagnosis* / Q000473:pathology*; D013737:Testis / Q000382:microbiology / Q000473:pathology; D014210:Treponema pallidum / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D015345:Oligonucleotide Probes; D012336:RNA, Ribosomal, 16S
keywords: Fluorescence in situ hybridization (FISH); Syphilis; Treponema pallidum subsp. pallidum