PMID- 28300642 OWN - NLM STAT- MEDLINE DCOM- 20180514 LR - 20180514 IS - 1095-8274 (Electronic) IS - 1075-9964 (Linking) VI - 47 DP - 2017 Oct TI - Distribution and phylogeny of Brachyspira spp. in human intestinal spirochetosis revealed by FISH and 16S rRNA-gene analysis. PG - 25-32 LID - S1075-9964(17)30055-0 [pii] LID - 10.1016/j.anaerobe.2017.03.012 [doi] AB - During six years as German National Consultant Laboratory for Spirochetes we investigated 149 intestinal biopsies from 91 patients, which were histopathologically diagnosed with human intestinal spirochetosis (HIS), using fluorescence in situ hybridization (FISH) combined with 16S rRNA gene PCR and sequencing. Aim of this study was to complement histopathological findings with FISH and PCR for definite diagnosis and species identification of the causative pathogens. HIS is characterized by colonization of the colonic mucosa of the human distal intestinal tract by Brachyspira spp. Microbiological diagnosis of HIS is not performed, because of the fastidious nature and slow growth of Brachyspira spp. in culture. In clinical practice, diagnosis of HIS relies solely on histopathology without differentiation of the spirochetes. We used a previously described FISH probe to detect and identify Brachyspira spp. in histological gut biopsies. FISH allowed rapid visualization and identification of Brachyspira spp. in 77 patients. In most cases, the bright FISH signal already allowed rapid localization of Brachyspira spp. at 400x magnification. By sequencing, 53 cases could be assigned to the B. aalborgi lineage including "B. ibaraki" and "B. hominis", and 23 cases to B. pilosicoli. One case showed mixed colonization. The cases reported here reaffirm all major HIS Brachyspira spp. clusters already described. However, the phylogenetic diversity seems to be even greater than previously reported. In 14 cases, we could not confirm HIS by either FISH or PCR, but found colonization of the epithelium by rods and cocci, indicating misdiagnosis by histopathology. FISH in combination with molecular identification by 16S rRNA gene sequencing has proved to be a valuable addition to histopathology. It provides definite diagnosis of HIS and allows insights into phylogeny and distribution of Brachyspira spp. HIS should be considered as a differential diagnosis in diarrhea of unknown origin, particularly in patients from risk groups (e.g. patients with colonic adenomas, inflammatory polyps, inflammatory bowel disease or HIV infection and in men who have sex with men). CI - Copyright (c) 2017 Elsevier Ltd. All rights reserved. FAU - Rojas, Pablo AU - Rojas P AD - Institute for Microbiology and Hygiene, Charite - Universitatsmedizin Berlin, Berlin, Germany. FAU - Petrich, Annett AU - Petrich A AD - Institute for Microbiology and Hygiene, Charite - Universitatsmedizin Berlin, Berlin, Germany. FAU - Schulze, Julia AU - Schulze J AD - Biofilmcenter, Deutsches Herzzentrum Berlin, Berlin, Germany. FAU - Wiessner, Alexandra AU - Wiessner A AD - Biofilmcenter, Deutsches Herzzentrum Berlin, Berlin, Germany. FAU - Loddenkemper, Christoph AU - Loddenkemper C AD - PathoTres Practice for Pathology, 12247 Berlin, Germany. FAU - Epple, Hans-Jorg AU - Epple HJ AD - Medical Clinic I, Gastroenterology, Rheumatology, Infectiology, Charite - Universitatsmedizin Berlin, Berlin, Germany. FAU - Sterlacci, William AU - Sterlacci W AD - Institute of Pathology, Klinikum Bayreuth GmbH, Bayreuth, Germany. FAU - Vieth, Michael AU - Vieth M AD - Institute of Pathology, Klinikum Bayreuth GmbH, Bayreuth, Germany. FAU - Kikhney, Judith AU - Kikhney J AD - Institute for Microbiology and Hygiene, Charite - Universitatsmedizin Berlin, Berlin, Germany; Biofilmcenter, Deutsches Herzzentrum Berlin, Berlin, Germany. FAU - Moter, Annette AU - Moter A AD - Biofilmcenter, Deutsches Herzzentrum Berlin, Berlin, Germany; Former German Consultant Laboratory for Treponema Identification, Charite - Universitatsmedizin Berlin, Berlin, Germany. Electronic address: moter@dhzb.de. LA - eng PT - Journal Article DEP - 20170312 PL - England TA - Anaerobe JT - Anaerobe JID - 9505216 RN - 0 (DNA, Bacterial) RN - 0 (DNA, Ribosomal) RN - 0 (RNA, Ribosomal, 16S) SB - IM MH - Adolescent MH - Adult MH - Aged MH - Aged, 80 and over MH - Brachyspira/*classification/genetics/*isolation & purification MH - Cluster Analysis MH - DNA, Bacterial/chemistry/genetics MH - DNA, Ribosomal/chemistry/genetics MH - Female MH - Genes, rRNA MH - *Genetic Variation MH - Gram-Negative Bacterial Infections/*microbiology MH - Humans MH - In Situ Hybridization, Fluorescence MH - Male MH - Middle Aged MH - Phylogeny MH - Polymerase Chain Reaction MH - RNA, Ribosomal, 16S/genetics MH - Sequence Analysis, DNA MH - Young Adult OTO - NOTNLM OT - 16S rRNA gene PCR OT - Brachyspira spp. OT - Fluorescence in situ hybridization (FISH) OT - Human intestinal spirochetosis (HIS) OT - Phylogeny EDAT- 2017/03/17 06:00 MHDA- 2018/05/15 06:00 CRDT- 2017/03/17 06:00 PHST- 2016/12/13 00:00 [received] PHST- 2017/03/09 00:00 [revised] PHST- 2017/03/10 00:00 [accepted] PHST- 2017/03/17 06:00 [pubmed] PHST- 2018/05/15 06:00 [medline] PHST- 2017/03/17 06:00 [entrez] AID - S1075-9964(17)30055-0 [pii] AID - 10.1016/j.anaerobe.2017.03.012 [doi] PST - ppublish SO - Anaerobe. 2017 Oct;47:25-32. doi: 10.1016/j.anaerobe.2017.03.012. Epub 2017 Mar 12.