title: In Situ Detection of Bacteria within Paraffin-embedded Tissues Using a Digoxin-labeled DNA Probe Targeting 16S rRNA.
abstract: The presence of bacteria within the pocket epithelium and underlying connective tissue in gingival biopsies from patients with periodontitis has been reported using various methods, including electron microscopy, immunohistochemistry or immunofluorescence using bacteria-specific antibodies, and fluorescent in situ hybridization (FISH) using a fluorescence-labeled oligonucleotide probe. Nevertheless, these methods are not widely used due to technical limitation or difficulties. Here a method to localize bacteria within paraffin-embedded tissues using DIG-labeled DNA probes has been introduced. The paraffin-embedded tissues are the most common form of biopsy tissues available from pathology banks. Bacteria can be detected either in a species-specific or universal manner. Bacterial signals are detected as either discrete forms (coccus, rod, fusiform, and hairy form) of bacteria or dispersed forms. The technique allows other histological information to be obtained: the epithelia, connective tissue, inflammatory infiltrates, and blood vessels are well distinguished. This method can be used to study the role of bacteria in various diseases, such as periodontitis, cancers, and inflammatory immune diseases.
authors.affiliation: Department of Oral Microbiology and Immunology, School of Dentistry and Dental Research Institute, Seoul National University.
authors.forename: Yun Sik
authors.initials: YS
authors.lastname: Choi
authors.affiliation: Department of Oral Microbiology and Immunology, School of Dentistry and Dental Research Institute, Seoul National University.
authors.forename: Yong Cheol
authors.initials: YC
authors.lastname: Kim
authors.affiliation: Department of Oral Microbiology and Immunology, School of Dentistry and Dental Research Institute, Seoul National University.
authors.forename: Keum Jin
authors.initials: KJ
authors.lastname: Baek
authors.affiliation: Department of Oral Microbiology and Immunology, School of Dentistry and Dental Research Institute, Seoul National University; youngnim@snu.ac.kr.
authors.forename: Youngnim
authors.initials: Y
authors.lastname: Choi
mesh_terms: D000818:Animals; D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D015342:DNA Probes / Q000737:chemistry* / Q000235:genetics; D004077:Digoxin / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry; D016480:Helicobacter pylori / Q000302:isolation & purification; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008010:Lichen Planus / Q000382:microbiology; D008175:Lung Neoplasms / Q000382:microbiology; D051379:Mice; D008807:Mice, Inbred BALB C; D016612:Paraffin Embedding / Q000379:methods; D016966:Porphyromonas gingivalis / Q000302:isolation & purification; D012336:RNA, Ribosomal, 16S / Q000032:analysis* / Q000235:genetics
chemical_list: D015342:DNA Probes; D005456:Fluorescent Dyes; D012336:RNA, Ribosomal, 16S; D004077:Digoxin