PMID- 15810941
OWN - NLM
STAT- MEDLINE
DCOM- 20050920
LR  - 20061115
IS  - 1083-4389 (Print)
IS  - 1083-4389 (Linking)
VI  - 10
IP  - 2
DP  - 2005 Apr
TI  - Visualization of Helicobacter species within the murine cecal mucosa using 
      specific fluorescence in situ hybridization.
PG  - 114-24
AB  - BACKGROUND: Members of the genus Helicobacter have been associated with colitis 
      development in a number of immunodeficient animal models. While it is known that 
      these organisms can initiate colitis development, the location and spatial 
      distribution of these bacteria within the intestinal tract is currently unknown. 
      In this study, we developed and optimized fluorescence in situ hybridization 
      (FISH) probes specifically for Helicobacter species. MATERIALS AND METHODS: Based 
      on 16S-RNA gene alignments, two probes specific for the entire family 
      Helicobacteraceae and two probes specific for Helicobacter ganmani and 
      Helicobacter hepaticus were designed. Evaluation of these probes was determined 
      using ATCC reference strains and cecum samples from ten IL-10 knockout mice. The 
      presence of Helicobacter species was determined using FISH and verified using 
      PCR-DGGE and microscopic examination of silver stained sections. RESULTS: 
      Analysis of the ATCC reference strains revealed that the probes HEL274/HEL717 
      were specific for the family Helicobacteraceae, while HEP642 was specific for H. 
      hepaticus and GAN1237 for H. ganmani. Using these probes, a pattern of spatial 
      localization of the two different Helicobacter species was observed in the cecum 
      tissues of IL-10 knockout mice. This consistently showed that H. ganmani was 
      localized to the lower regions and H. hepaticus to the mid-upper regions of the 
      crypts. CONCLUSION: We have developed FISH probes specific for the family 
      Helicobacteraceae as well as two individual Helicobacter species. This study will 
      allow the future use of the FISH to better understand host-pathogen interactions 
      in vitro.
FAU - Chan, Vivian
AU  - Chan V
AD  - School of Biotechnology and Biomolecular Sciences, The University of New South 
      Wales, Sydney, NSW 2052, Australia.
FAU - Crocetti, Gregory
AU  - Crocetti G
FAU - Grehan, Martin
AU  - Grehan M
FAU - Zhang, Li
AU  - Zhang L
FAU - Danon, Stephen
AU  - Danon S
FAU - Lee, Adrian
AU  - Lee A
FAU - Mitchell, Hazel
AU  - Mitchell H
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Helicobacter
JT  - Helicobacter
JID - 9605411
RN  - 0 (Oligonucleotide Probes)
RN  - 0 (RNA, Bacterial)
RN  - 0 (RNA, Ribosomal, 16S)
RN  - 130068-27-8 (Interleukin-10)
SB  - IM
MH  - Animals
MH  - Cecum/*microbiology/pathology
MH  - Helicobacter/genetics/*isolation & purification
MH  - Helicobacter Infections/*diagnosis/microbiology/pathology
MH  - Helicobacter hepaticus/genetics/isolation & purification
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Interleukin-10/deficiency/genetics
MH  - Intestinal Mucosa/*microbiology
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Mice, Knockout
MH  - Oligonucleotide Probes
MH  - RNA, Bacterial/genetics
MH  - RNA, Ribosomal, 16S/genetics
EDAT- 2005/04/07 09:00
MHDA- 2005/09/21 09:00
CRDT- 2005/04/07 09:00
PHST- 2005/04/07 09:00 [pubmed]
PHST- 2005/09/21 09:00 [medline]
PHST- 2005/04/07 09:00 [entrez]
AID - HEL298 [pii]
AID - 10.1111/j.1523-5378.2005.00298.x [doi]
PST - ppublish
SO  - Helicobacter. 2005 Apr;10(2):114-24. doi: 10.1111/j.1523-5378.2005.00298.x.