PMID- 20453122
OWN - NLM
STAT- MEDLINE
DCOM- 20100907
LR  - 20240318
IS  - 1098-5336 (Electronic)
IS  - 0099-2240 (Print)
IS  - 0099-2240 (Linking)
VI  - 76
IP  - 13
DP  - 2010 Jul
TI  - Fluorescence in situ hybridization method using a peptide nucleic acid probe for 
      identification of Salmonella spp. in a broad spectrum of samples.
PG  - 4476-85
LID - 10.1128/AEM.01678-09 [doi]
AB  - A fluorescence in situ hybridization (FISH) method for the rapid detection of 
      Salmonella spp. using a novel peptide nucleic acid (PNA) probe was developed. The 
      probe theoretical specificity and sensitivity were both 100%. The PNA-FISH method 
      was optimized, and laboratory testing on representative strains from the 
      Salmonella genus subspecies and several related bacterial species confirmed the 
      predicted theoretical values of specificity and sensitivity. The PNA-FISH method 
      has been successfully adapted to detect cells in suspension and is hence able to 
      be employed for the detection of this bacterium in blood, feces, water, and 
      powdered infant formula (PIF). The blood and PIF samples were artificially 
      contaminated with decreasing pathogen concentrations. After the use of an 
      enrichment step, the PNA-FISH method was able to detect 1 CFU per 10 ml of blood 
      (5 x 10(9) +/- 5 x 10(8) CFU/ml after an overnight enrichment step) and also 1 
      CFU per 10 g of PIF (2 x 10(7) +/- 5 x 10(6) CFU/ml after an 8-h enrichment 
      step). The feces and water samples were also enriched according to the 
      corresponding International Organization for Standardization methods, and results 
      showed that the PNA-FISH method was able to detect Salmonella immediately after 
      the first enrichment step was conducted. Moreover, the probe was able to 
      discriminate the bacterium in a mixed microbial population in feces and water by 
      counter-staining with 4',6-diamidino-2-phenylindole (DAPI). This new method is 
      applicable to a broad spectrum of samples and takes less than 20 h to obtain a 
      diagnosis, except for PIF samples, where the analysis takes less than 12 h. This 
      procedure may be used for food processing and municipal water control and also in 
      clinical settings, representing an improved alternative to culture-based 
      techniques and to the existing Salmonella PNA probe, Sal23S10, which presents a 
      lower specificity.
FAU - Almeida, C
AU  - Almeida C
AD  - Centro de Engenharia Biologica, Universidade do Minho, 4710-057 Braga, Portugal.
FAU - Azevedo, N F
AU  - Azevedo NF
FAU - Fernandes, R M
AU  - Fernandes RM
FAU - Keevil, C W
AU  - Keevil CW
FAU - Vieira, M J
AU  - Vieira MJ
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20100507
PL  - United States
TA  - Appl Environ Microbiol
JT  - Applied and environmental microbiology
JID - 7605801
RN  - 0 (Oligonucleotide Probes)
RN  - 0 (Peptide Nucleic Acids)
SB  - IM
MH  - Blood/microbiology
MH  - *Environmental Microbiology
MH  - Feces/microbiology
MH  - *Food Microbiology
MH  - Fresh Water/microbiology
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Infant Formula
MH  - Infant, Newborn
MH  - Oligonucleotide Probes/*genetics
MH  - Peptide Nucleic Acids/*genetics
MH  - *Salmonella/classification/genetics/isolation & purification
MH  - Salmonella Infections/*microbiology
MH  - Sensitivity and Specificity
MH  - Water Supply
PMC - PMC2897454
EDAT- 2010/05/11 06:00
MHDA- 2010/09/08 06:00
PMCR- 2011/01/01
CRDT- 2010/05/11 06:00
PHST- 2010/05/11 06:00 [entrez]
PHST- 2010/05/11 06:00 [pubmed]
PHST- 2010/09/08 06:00 [medline]
PHST- 2011/01/01 00:00 [pmc-release]
AID - AEM.01678-09 [pii]
AID - 1678-09 [pii]
AID - 10.1128/AEM.01678-09 [doi]
PST - ppublish
SO  - Appl Environ Microbiol. 2010 Jul;76(13):4476-85. doi: 10.1128/AEM.01678-09. Epub 
      2010 May 7.