PMID- 16382905
OWN - NLM
STAT- MEDLINE
DCOM- 20060118
LR  - 20180607
IS  - 0736-6205 (Print)
IS  - 0736-6205 (Linking)
VI  - 39
IP  - 6
DP  - 2005 Dec
TI  - Laser capture microdissection of bacterial cells targeted by fluorescence in situ 
      hybridization.
PG  - 864-8
AB  - Direct cultivation-independent sequence retrieval of unidentified bacteria from 
      histological tissue sections has been limited by the difficulty of selectively 
      isolating specific bacteria from a complex environment. Here, a new DNA isolation 
      approach is presented for prokaryotic cells. By this method, a potentially 
      pathogenic strain of the genus Brachyspira from formalin-fixed human colonic 
      biopsies were visualized by fluorescence in situ hybridization (FISH) with a 16S 
      rRNA-targeting oligonucleotide probe, followed by laser capture microdissection 
      (LCM) of the targeted cells. Direct 16S rRNA gene PCR was performed from the 
      dissected microcolonies, and the subsequent DNA sequence analysis identified the 
      dissected bacterial cells as belonging to the Brachyspira aalborgi cluster 1. The 
      advantage of this technique is the ability to combine the histological 
      recognition of the specific bacteria within the tissue with molecular analysis of 
      16S rRNA gene or other genes of interest. This method is widely applicable for 
      the identification of noncultivable bacteria and their gene pool from 
      formalin-fixed paraffin-embedded tissue samples.
FAU - Klitgaard, Kirstine
AU  - Klitgaard K
AD  - Danish Institute for Food and Veterinary Research, Copenhagen, Denmark.
FAU - Molbak, Lars
AU  - Molbak L
FAU - Jensen, Tim K
AU  - Jensen TK
FAU - Lindboe, Christian Fredrik
AU  - Lindboe CF
FAU - Boye, Mette
AU  - Boye M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Biotechniques
JT  - BioTechniques
JID - 8306785
RN  - 0 (DNA, Bacterial)
RN  - 0 (DNA, Ribosomal)
RN  - 0 (RNA, Ribosomal, 16S)
SB  - IM
MH  - Brachyspira/genetics/*isolation & purification
MH  - DNA, Bacterial/*analysis
MH  - DNA, Ribosomal/analysis
MH  - Humans
MH  - *In Situ Hybridization, Fluorescence
MH  - Lasers
MH  - Microdissection/*methods
MH  - RNA, Ribosomal, 16S/genetics
EDAT- 2005/12/31 09:00
MHDA- 2006/01/19 09:00
CRDT- 2005/12/31 09:00
PHST- 2005/12/31 09:00 [pubmed]
PHST- 2006/01/19 09:00 [medline]
PHST- 2005/12/31 09:00 [entrez]
AID - 000112024 [pii]
AID - 10.2144/000112024 [doi]
PST - ppublish
SO  - Biotechniques. 2005 Dec;39(6):864-8. doi: 10.2144/000112024.