PMID- 26813295
OWN - NLM
STAT- MEDLINE
DCOM- 20160921
LR  - 20181202
IS  - 1029-2454 (Electronic)
IS  - 0892-7014 (Linking)
VI  - 32
IP  - 2
DP  - 2016
TI  - Discrimination of bacteriophage infected cells using locked nucleic acid 
      fluorescent in situ hybridization (LNA-FISH).
PG  - 179-90
LID - 10.1080/08927014.2015.1131821 [doi]
AB  - Bacteriophage-host interaction studies in biofilm structures are still 
      challenging due to the technical limitations of traditional methods. The aim of 
      this study was to provide a direct fluorescence in situ hybridization (FISH) 
      method based on locked nucleic acid (LNA) probes, which targets the phage 
      replication phase, allowing the study of population dynamics during infection. 
      Bacteriophages specific for two biofilm-forming bacteria, Pseudomonas aeruginosa 
      and Acinetobacter, were selected. Four LNA probes were designed and optimized for 
      phage-specific detection and for bacterial counterstaining. To validate the 
      method, LNA-FISH counts were compared with the traditional plaque forming unit 
      (PFU) technique. To visualize the progression of phage infection within a 
      biofilm, colony-biofilms were formed and infected with bacteriophages. A good 
      correlation (r = 0.707) was observed between LNA-FISH and PFU techniques. In 
      biofilm structures, LNA-FISH provided a good discrimination of the infected cells 
      and also allowed the assessment of the spatial distribution of infected and 
      non-infected populations.
FAU - Vilas Boas, Diana
AU  - Vilas Boas D
AD  - b Laboratory for Process, Environment, Biotechnology and Energy Engineering 
      (LEPABE), Department of Chemical Engineering, Faculty of Engineering , University 
      of Porto , Porto , Portugal.
FAU - Almeida, Carina
AU  - Almeida C
AD  - a LIBRO -Laboratorio de Investigacao em Biofilmes Rosario Oliveira , Centre of 
      Biological Engineering, University of Minho , Braga , Portugal.
AD  - b Laboratory for Process, Environment, Biotechnology and Energy Engineering 
      (LEPABE), Department of Chemical Engineering, Faculty of Engineering , University 
      of Porto , Porto , Portugal.
FAU - Sillankorva, Sanna
AU  - Sillankorva S
AD  - a LIBRO -Laboratorio de Investigacao em Biofilmes Rosario Oliveira , Centre of 
      Biological Engineering, University of Minho , Braga , Portugal.
FAU - Nicolau, Ana
AU  - Nicolau A
AD  - b Laboratory for Process, Environment, Biotechnology and Energy Engineering 
      (LEPABE), Department of Chemical Engineering, Faculty of Engineering , University 
      of Porto , Porto , Portugal.
FAU - Azeredo, Joana
AU  - Azeredo J
AD  - b Laboratory for Process, Environment, Biotechnology and Energy Engineering 
      (LEPABE), Department of Chemical Engineering, Faculty of Engineering , University 
      of Porto , Porto , Portugal.
FAU - Azevedo, Nuno F
AU  - Azevedo NF
AD  - b Laboratory for Process, Environment, Biotechnology and Energy Engineering 
      (LEPABE), Department of Chemical Engineering, Faculty of Engineering , University 
      of Porto , Porto , Portugal.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Biofouling
JT  - Biofouling
JID - 9200331
RN  - 0 (Oligonucleotides)
RN  - 0 (locked nucleic acid)
SB  - IM
EIN - Biofouling. 2016;32(2):ii. PMID: 26901589
MH  - Acinetobacter baumannii/*virology
MH  - Bacteriophages/*physiology
MH  - Biofilms/*growth & development
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Oligonucleotides/metabolism
MH  - Pseudomonas aeruginosa/*virology
MH  - Reproducibility of Results
OTO - NOTNLM
OT  - Acinetobacter
OT  - Pseudomonas aeruginosa
OT  - bacteriophage
OT  - biofilms
OT  - phage infection, LNA-FISH
EDAT- 2016/01/28 06:00
MHDA- 2016/09/23 06:00
CRDT- 2016/01/28 06:00
PHST- 2016/01/28 06:00 [entrez]
PHST- 2016/01/28 06:00 [pubmed]
PHST- 2016/09/23 06:00 [medline]
AID - 10.1080/08927014.2015.1131821 [doi]
PST - ppublish
SO  - Biofouling. 2016;32(2):179-90. doi: 10.1080/08927014.2015.1131821.