PMID- 25840566
OWN - NLM
STAT- MEDLINE
DCOM- 20151228
LR  - 20150416
IS  - 1432-0614 (Electronic)
IS  - 0175-7598 (Linking)
VI  - 99
IP  - 9
DP  - 2015 May
TI  - Mismatch discrimination in fluorescent in situ hybridization using different 
      types of nucleic acids.
PG  - 3961-9
LID - 10.1007/s00253-015-6389-4 [doi]
AB  - In the past few years, several researchers have focused their attention on 
      nucleic acid mimics due to the increasing necessity of developing a more robust 
      recognition of DNA or RNA sequences. Fluorescence in situ hybridization (FISH) is 
      an example of a method where the use of these novel nucleic acid monomers might 
      be crucial to the success of the analysis. To achieve the expected accuracy in 
      detection, FISH probes should have high binding affinity towards their 
      complementary strands and discriminate effectively the noncomplementary strands. 
      In this study, we investigate the effect of different chemical modifications in 
      fluorescent probes on their ability to successfully detect the complementary 
      target and discriminate the mismatched base pairs by FISH. To our knowledge, this 
      paper presents the first study where this analysis is performed with different 
      types of FISH probes directly in biological targets, Helicobacter pylori and 
      Helicobacter acinonychis. This is also the first study where unlocked nucleic 
      acids (UNA) were used as chemistry modification in oligonucleotides for FISH 
      methodologies. The effectiveness in detecting the specific target and in mismatch 
      discrimination appears to be improved using locked nucleic acids 
      (LNA)/2'-O-methyl RNA (2'OMe) or peptide nucleic acid (PNA) in comparison to 
      LNA/DNA, LNA/UNA, or DNA probes. Further, the use of LNA modifications together 
      with 2'OMe monomers allowed the use of shorter fluorescent probes and increased 
      the range of hybridization temperatures at which FISH would work.
FAU - Fontenete, Silvia
AU  - Fontenete S
FAU - Barros, Joana
AU  - Barros J
FAU - Madureira, Pedro
AU  - Madureira P
FAU - Figueiredo, Ceu
AU  - Figueiredo C
FAU - Wengel, Jesper
AU  - Wengel J
FAU - Azevedo, Nuno Filipe
AU  - Azevedo NF
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20150121
PL  - Germany
TA  - Appl Microbiol Biotechnol
JT  - Applied microbiology and biotechnology
JID - 8406612
RN  - 0 (Nucleic Acids)
RN  - 0 (Oligonucleotide Probes)
SB  - IM
EIN - Appl Microbiol Biotechnol. 2015 May;99(9):4117. Silvia, Fontenete [corrected to 
      Fontenete, Silvia]; Joana, Barros [corrected to Barros, Joana]; Pedro, Madureira 
      [corrected to Madureira, Pedro]; Ceu, Figueiredo [corrected to Figueiredo, Ceu]; 
      Jesper, Wengel [corrected to Wengel, Jesper]; Filipe, Azeved. PMID: 25846338
MH  - *Base Pair Mismatch
MH  - Helicobacter pylori/genetics
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Molecular Diagnostic Techniques/*methods
MH  - Nucleic Acids/*genetics
MH  - Oligonucleotide Probes/genetics
MH  - Sensitivity and Specificity
MH  - Temperature
EDAT- 2015/04/05 06:00
MHDA- 2015/12/29 06:00
CRDT- 2015/04/05 06:00
PHST- 2014/08/20 00:00 [received]
PHST- 2014/11/08 00:00 [accepted]
PHST- 2014/11/07 00:00 [revised]
PHST- 2015/04/05 06:00 [entrez]
PHST- 2015/04/05 06:00 [pubmed]
PHST- 2015/12/29 06:00 [medline]
AID - 10.1007/s00253-015-6389-4 [doi]
PST - ppublish
SO  - Appl Microbiol Biotechnol. 2015 May;99(9):3961-9. doi: 10.1007/s00253-015-6389-4. 
      Epub 2015 Jan 21.