PMID- 12724371
OWN - NLM
STAT- MEDLINE
DCOM- 20030731
LR  - 20200826
IS  - 1350-0872 (Print)
IS  - 1350-0872 (Linking)
VI  - 149
IP  - Pt 5
DP  - 2003 May
TI  - Fluorescence in situ hybridization (FISH) for direct visualization of bacteria in 
      periapical lesions of asymptomatic root-filled teeth.
PG  - 1095-1102
LID - 10.1099/mic.0.26077-0 [doi]
AB  - Whether micro-organisms can live in periapical endodontic lesions of asymptomatic 
      teeth is under debate. The aim of the present study was to visualize and identify 
      micro-organisms within periapical lesions directly, using fluorescence in situ 
      hybridization (FISH) in combination with epifluorescence and confocal laser 
      scanning microscopy (CLSM). Thirty-nine periapical lesions were surgically 
      removed, fixed, embedded in cold polymerizing resin and sectioned. The probe EUB 
      338, specific for the domain Bacteria, was used together with a number of 
      species-specific 16S rRNA-directed oligonucleotide probes to identify bacteria. 
      To control non-specific binding of EUB 338, probe NON 338 was used. 
      Alternatively, DAPI (4',6'-diamidino-2-phenylindole) staining was applied to 
      record prokaryotic and eukaryotic DNA in the specimens. Hybridization with NON 
      338 gave no signals despite background fluorescence of the tissue. The 
      eubacterial probe showed bacteria of different morphotypes in 50 % of the 
      lesions. Rods, spirochaetes and cocci were spread out in areas of the tissue 
      while other parts seemed bacteria-free. Bacteria were also seen to co-aggregate 
      inside the tissue, forming microcolonies. Porphyromonas gingivalis, Prevotella 
      intermedia, Tannerella forsythensis and treponemes of phylogenetic Group I were 
      detected with specific probes. In addition, colonies with Streptococcus spp. were 
      seen in some lesions. A number of morphotypes occurred that could not be 
      identified with the specific probes used, indicating the presence of additional 
      bacterial species. CLSM confirmed that bacteria were located in different layers 
      of the tissue. Accordingly, the FISH technique demonstrated mixed consortia of 
      bacteria consisting of rods, spirochaetes and cocci in asymptomatic periapical 
      lesions of root-filled teeth.
FAU - Sunde, Pia T
AU  - Sunde PT
AD  - Institute of Oral Biology, Dental Faculty, University of Oslo, PB 1052 Blindern, 
      0316 Oslo, Norway.
FAU - Olsen, Ingar
AU  - Olsen I
AD  - Institute of Oral Biology, Dental Faculty, University of Oslo, PB 1052 Blindern, 
      0316 Oslo, Norway.
FAU - Gobel, Ulf B
AU  - Gobel UB
AD  - Institut fur Mikrobiologie und Hygiene, Universitatsklinikum Charite, 
      Humboldt-Universitat zu Berlin, Dorotheenstrasse 96, D-10117 Berlin, Germany.
FAU - Theegarten, Dirk
AU  - Theegarten D
AD  - Abteilung fur Pathologie, Ruhr-Universitat Bochum, Universitatsstrasse 150, 
      D-44780 Bochum, Germany.
FAU - Winter, Sascha
AU  - Winter S
AD  - Universitatsklinik fur Mund-, Kiefer- und Plastische Gesichtschirurgie, 
      Knappschafts-Krankenhaus Bochum-Langendreer, In der Schornau 23-25, D-44892 
      Bochum, Germany.
FAU - Debelian, Gilberto J
AU  - Debelian GJ
AD  - Institute of Oral Biology, Dental Faculty, University of Oslo, PB 1052 Blindern, 
      0316 Oslo, Norway.
FAU - Tronstad, Leif
AU  - Tronstad L
AD  - Institute of Oral Biology, Dental Faculty, University of Oslo, PB 1052 Blindern, 
      0316 Oslo, Norway.
FAU - Moter, Annette
AU  - Moter A
AD  - Institut fur Mikrobiologie und Hygiene, Universitatsklinikum Charite, 
      Humboldt-Universitat zu Berlin, Dorotheenstrasse 96, D-10117 Berlin, Germany.
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PL  - England
TA  - Microbiology (Reading)
JT  - Microbiology (Reading, England)
JID - 9430468
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Oligonucleotide Probes)
RN  - 0 (RNA, Ribosomal, 16S)
SB  - IM
MH  - Bacteria/*classification/genetics/*isolation & purification
MH  - Bacterial Infections/microbiology
MH  - Colony Count, Microbial
MH  - Dental Pulp Cavity/*microbiology
MH  - Fluorescent Dyes
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Male
MH  - Microscopy, Confocal
MH  - Microscopy, Fluorescence
MH  - Middle Aged
MH  - Oligonucleotide Probes
MH  - Periapical Periodontitis/*microbiology
MH  - RNA, Ribosomal, 16S/genetics
MH  - Species Specificity
MH  - Tooth/*microbiology
MH  - Tooth Root/microbiology
EDAT- 2003/05/02 05:00
MHDA- 2003/08/02 05:00
CRDT- 2003/05/02 05:00
PHST- 2003/05/02 05:00 [pubmed]
PHST- 2003/08/02 05:00 [medline]
PHST- 2003/05/02 05:00 [entrez]
AID - 10.1099/mic.0.26077-0 [doi]
PST - ppublish
SO  - Microbiology (Reading). 2003 May;149(Pt 5):1095-1102. doi: 10.1099/mic.0.26077-0.