Application of locked nucleic acid-based probes in fluorescence in situ hybridization.
Identifieur interne : 001373 ( Main/Exploration ); précédent : 001372; suivant : 001374Application of locked nucleic acid-based probes in fluorescence in situ hybridization.
Auteurs : Sílvia Fontenete [Portugal] ; Daniel Carvalho [Portugal] ; Nuno Guimarães [Portugal] ; Pedro Madureira [Portugal] ; Céu Figueiredo [Portugal] ; Jesper Wengel [Danemark] ; Nuno Filipe Azevedo [Portugal]Source :
- Applied microbiology and biotechnology [ 1432-0614 ] ; 2016.
Descripteurs français
- KwdFr :
- Helicobacter pylori (génétique), Helicobacter pylori (isolement et purification), Helicobacter pylori (métabolisme), Oligonucléotides (génétique), Oligonucléotides (métabolisme), Protéines bactériennes (génétique), Protéines bactériennes (métabolisme), Sondes oligonucléotidiques (génétique), Sondes oligonucléotidiques (métabolisme), Technique FISH (), Technique FISH (instrumentation).
- MESH :
- génétique : Helicobacter pylori, Oligonucléotides, Protéines bactériennes, Sondes oligonucléotidiques.
- isolement et purification : Helicobacter pylori.
- métabolisme : Helicobacter pylori, Oligonucléotides, Protéines bactériennes, Sondes oligonucléotidiques, Technique FISH.
- Technique FISH.
English descriptors
- KwdEn :
- Bacterial Proteins (genetics), Bacterial Proteins (metabolism), Helicobacter pylori (genetics), Helicobacter pylori (isolation & purification), Helicobacter pylori (metabolism), In Situ Hybridization, Fluorescence (instrumentation), In Situ Hybridization, Fluorescence (methods), Oligonucleotide Probes (genetics), Oligonucleotide Probes (metabolism), Oligonucleotides (genetics), Oligonucleotides (metabolism).
- MESH :
- chemical , genetics : Bacterial Proteins, Oligonucleotide Probes, Oligonucleotides.
- chemical , metabolism : Bacterial Proteins, Oligonucleotide Probes, Oligonucleotides.
- genetics : Helicobacter pylori.
- instrumentation : In Situ Hybridization, Fluorescence.
- isolation & purification : Helicobacter pylori.
- metabolism : Helicobacter pylori.
- methods : In Situ Hybridization, Fluorescence.
Abstract
Fluorescence in situ hybridization (FISH) employing nucleic acid mimics as probes is becoming an emerging molecular tool in the microbiology area for the detection and visualization of microorganisms. However, the impact that locked nucleic acid (LNA) and 2'-O-methyl (2'-OMe) RNA modifications have on the probe that is targeting microorganisms is unknown. In this study, the melting and hybridization efficiency properties of 18 different probes in regards to their use in FISH for the detection of the 16S rRNA of Helicobacter pylori were compared. For the same sequence and target, probe length and the type of nucleic acid mimics used as mixmers in LNA-based probes strongly influence the efficiency of detection. LNA probes with 10 to 15 mers showed the highest efficiency. Additionally, the combination of 2'-OMe RNA with LNA allowed an increase on the fluorescence intensities of the probes. Overall, these results have significant implications for the design and applications of LNA probes for the detection of microorganisms.
DOI: 10.1007/s00253-016-7429-4
PubMed: 26969040
Affiliations:
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xml:lang="en"><term>Helicobacter pylori</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Helicobacter pylori</term><term>Oligonucléotides</term><term>Protéines bactériennes</term><term>Sondes oligonucléotidiques</term></keywords><keywords scheme="MESH" qualifier="instrumentation" xml:lang="en"><term>In Situ Hybridization, Fluorescence</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Helicobacter pylori</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Helicobacter pylori</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Helicobacter pylori</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>In Situ Hybridization, Fluorescence</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Helicobacter pylori</term><term>Oligonucléotides</term><term>Protéines 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However, the impact that locked nucleic acid (LNA) and 2'-O-methyl (2'-OMe) RNA modifications have on the probe that is targeting microorganisms is unknown. In this study, the melting and hybridization efficiency properties of 18 different probes in regards to their use in FISH for the detection of the 16S rRNA of Helicobacter pylori were compared. For the same sequence and target, probe length and the type of nucleic acid mimics used as mixmers in LNA-based probes strongly influence the efficiency of detection. LNA probes with 10 to 15 mers showed the highest efficiency. Additionally, the combination of 2'-OMe RNA with LNA allowed an increase on the fluorescence intensities of the probes. Overall, these results have significant implications for the design and applications of LNA probes for the detection of microorganisms. </div></front></TEI><affiliations><list><country><li>Danemark</li><li>Portugal</li></country></list><tree><country name="Portugal"><noRegion><name sortKey="Fontenete, Silvia" sort="Fontenete, Silvia" uniqKey="Fontenete S" first="Sílvia" last="Fontenete">Sílvia Fontenete</name></noRegion><name sortKey="Azevedo, Nuno Filipe" sort="Azevedo, Nuno Filipe" uniqKey="Azevedo N" first="Nuno Filipe" last="Azevedo">Nuno Filipe Azevedo</name><name sortKey="Carvalho, Daniel" sort="Carvalho, Daniel" uniqKey="Carvalho D" first="Daniel" last="Carvalho">Daniel Carvalho</name><name sortKey="Figueiredo, Ceu" sort="Figueiredo, Ceu" uniqKey="Figueiredo C" first="Céu" last="Figueiredo">Céu Figueiredo</name><name sortKey="Guimaraes, Nuno" sort="Guimaraes, Nuno" uniqKey="Guimaraes N" first="Nuno" last="Guimarães">Nuno Guimarães</name><name sortKey="Madureira, Pedro" sort="Madureira, Pedro" uniqKey="Madureira P" first="Pedro" last="Madureira">Pedro Madureira</name></country><country name="Danemark"><noRegion><name sortKey="Wengel, Jesper" sort="Wengel, Jesper" uniqKey="Wengel J" first="Jesper" last="Wengel">Jesper Wengel</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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