PMID- 24278398
OWN - NLM
STAT- MEDLINE
DCOM- 20150508
LR  - 20211021
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 8
IP  - 11
DP  - 2013
TI  - Hybridization-based detection of Helicobacter pylori at human body temperature 
      using advanced locked nucleic acid (LNA) probes.
PG  - e81230
LID - 10.1371/journal.pone.0081230 [doi]
LID - e81230
AB  - The understanding of the human microbiome and its influence upon human life has 
      long been a subject of study. Hence, methods that allow the direct detection and 
      visualization of microorganisms and microbial consortia (e.g. biofilms) within 
      the human body would be invaluable. In here, we assessed the possibility of 
      developing a variant of fluorescence in situ hybridization (FISH), named 
      fluorescence in vivo hybridization (FIVH), for the detection of Helicobacter 
      pylori. Using oligonucleotide variations comprising locked nucleic acids (LNA) 
      and 2'-O-methyl RNAs (2'OMe) with two types of backbone linkages (phosphate or 
      phosphorothioate), we were able to successfully identify two probes that 
      hybridize at 37  degrees C with high specificity and sensitivity for H. pylori, both in 
      pure cultures and in gastric biopsies. Furthermore, the use of this type of 
      probes implied that toxic compounds typically used in FISH were either found to 
      be unnecessary or could be replaced by a non-toxic substitute. We show here for 
      the first time that the use of advanced LNA probes in FIVH conditions provides an 
      accurate, simple and fast method for H. pylori detection and location, which 
      could be used in the future for potential in vivo applications either for this 
      microorganism or for others.
FAU - Fontenete, Silvia
AU  - Fontenete S
AD  - LEPABE, Laboratory for Process Engineering, Environment, Biotechnology and 
      Energy, Department of Chemical Engineering, Faculty of Engineering, University of 
      Porto, Porto, Portugal ; IPATIMUP, Institute of Molecular Pathology and 
      Immunology of the University of Porto, Porto, Portugal ; Nucleic Acid Center, 
      Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, 
      Odense M, Denmark ; ICBAS, Institute of Biomedical Sciences Abel Salazar, 
      University of Porto, Porto, Portugal.
FAU - Guimaraes, Nuno
AU  - Guimaraes N
FAU - Leite, Marina
AU  - Leite M
FAU - Figueiredo, Ceu
AU  - Figueiredo C
FAU - Wengel, Jesper
AU  - Wengel J
FAU - Filipe Azevedo, Nuno
AU  - Filipe Azevedo N
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20131122
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Nucleic Acid Probes)
SB  - IM
MH  - Flow Cytometry
MH  - Helicobacter Infections/diagnosis
MH  - Helicobacter pylori/*genetics
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Microscopy, Fluorescence
MH  - Nucleic Acid Hybridization
MH  - Nucleic Acid Probes
MH  - Stomach/microbiology
PMC - PMC3838382
COIS- Competing Interests: Jesper Wengel is cofounder of RiboTask ApS which offer 
      LNA/2'-OMe-RNA probes for RNA targeting. Nuno Filipe Azevedo is cofounder of 
      Biomode SA which develops molecular methods for the rapid detection of 
      microorganisms. This does not alter the authors' adherence to all the PLOS ONE 
      policies on sharing data and materials.
EDAT- 2013/11/28 06:00
MHDA- 2015/05/09 06:00
PMCR- 2013/11/22
CRDT- 2013/11/27 06:00
PHST- 2013/04/11 00:00 [received]
PHST- 2013/10/10 00:00 [accepted]
PHST- 2013/11/27 06:00 [entrez]
PHST- 2013/11/28 06:00 [pubmed]
PHST- 2015/05/09 06:00 [medline]
PHST- 2013/11/22 00:00 [pmc-release]
AID - PONE-D-13-15277 [pii]
AID - 10.1371/journal.pone.0081230 [doi]
PST - epublish
SO  - PLoS One. 2013 Nov 22;8(11):e81230. doi: 10.1371/journal.pone.0081230. 
      eCollection 2013.