PMID- 16151137
OWN - NLM
STAT- MEDLINE
DCOM- 20051101
LR  - 20191210
IS  - 0099-2240 (Print)
IS  - 1098-5336 (Electronic)
IS  - 0099-2240 (Linking)
VI  - 71
IP  - 9
DP  - 2005 Sep
TI  - Design and evaluation of 16S rRNA-targeted peptide nucleic acid probes for 
      whole-cell detection of members of the genus Listeria.
PG  - 5451-7
AB  - Six fluorescein-labeled peptide nucleic acid oligomers targeting 
      Listeria-specific sequences on the 16S ribosomal subunit were evaluated for their 
      abilities to hybridize to whole cells by fluorescence in situ hybridization 
      (FISH). Four of these probes yielded weak or no fluorescent signals after 
      hybridization and were not investigated further. The remaining two 
      FISH-compatible probes, LisUn-3 and LisUn-11, were evaluated for their 
      reactivities against 22 Listeria strains and 17 other bacterial strains belonging 
      to 10 closely related genera. Hybridization with BacUni-1, a domain-specific 
      eubacterial probe, was used as a positive control for target accessibility in 
      both Listeria spp. and nontarget cells. RNase T1 treatment of select cell types 
      was used to confirm that positive fluorescence responses were rRNA dependent and 
      to examine the extent of nonspecific staining of nontarget cells. Both LisUn-3 
      and LisUn-11 yielded rapid, bright, and genus-specific hybridizations at probe 
      concentrations of approximately 100 pmol ml(-1). LisUn-11 was the brightest probe 
      and stained all six Listeria species. LisUn-3 hybridized with all Listeria spp. 
      except for L. grayi, for which it had two mismatched bases. A simple ethanolic 
      fixation yielded superior results with Listeria spp. compared to fixation in 10% 
      buffered formalin and was applicable to all cell types studied. This study 
      highlights the advantages of peptide nucleic acid probes for FISH-based detection 
      of gram-positive bacteria and provides new tools for the rapid detection of 
      Listeria spp. These probes may be useful for the routine monitoring of food 
      production environments in support of efforts to control L. monocytogenes.
FAU - Brehm-Stecher, Byron F
AU  - Brehm-Stecher BF
AD  - Department of Food Science and Human Nutrition, 2312 Food Sciences Building, Iowa 
      State University, Ames, IA 50011, USA. byron@iastate.edu
FAU - Hyldig-Nielsen, Jens J
AU  - Hyldig-Nielsen JJ
FAU - Johnson, Eric A
AU  - Johnson EA
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Appl Environ Microbiol
JT  - Applied and environmental microbiology
JID - 7605801
RN  - 0 (Nucleic Acid Probes)
RN  - 0 (Peptide Nucleic Acids)
RN  - 0 (RNA, Ribosomal, 16S)
SB  - IM
MH  - Base Sequence
MH  - Flow Cytometry
MH  - Genes, rRNA
MH  - Humans
MH  - *In Situ Hybridization, Fluorescence
MH  - Listeria/classification/cytology/genetics/*isolation & purification
MH  - Molecular Sequence Data
MH  - Nucleic Acid Probes/*genetics
MH  - Peptide Nucleic Acids/*genetics
MH  - RNA, Ribosomal, 16S/*genetics
PMC - PMC1214657
EDAT- 2005/09/10 09:00
MHDA- 2005/11/03 09:00
PMCR- 2005/09/01
CRDT- 2005/09/10 09:00
PHST- 2005/09/10 09:00 [pubmed]
PHST- 2005/11/03 09:00 [medline]
PHST- 2005/09/10 09:00 [entrez]
PHST- 2005/09/01 00:00 [pmc-release]
AID - 71/9/5451 [pii]
AID - 2293-04 [pii]
AID - 10.1128/AEM.71.9.5451-5457.2005 [doi]
PST - ppublish
SO  - Appl Environ Microbiol. 2005 Sep;71(9):5451-7. doi: 
      10.1128/AEM.71.9.5451-5457.2005.