Mismatch discrimination in fluorescent in situ hybridization using different types of nucleic acids

Verfasser / Beitragende:
[Fontenete Silvia, Barros Joana, Madureira Pedro, Figueiredo Céu, Wengel Jesper, Azevedo Filipe]
Ort, Verlag, Jahr:
2015
Enthalten in:
Applied Microbiology and Biotechnology, 99/9(2015-05-01), 3961-3969
Format:
Artikel (online)
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024 7 0 |a 10.1007/s00253-015-6389-4  |2 doi 
035 |a (NATIONALLICENCE)springer-10.1007/s00253-015-6389-4 
245 0 0 |a Mismatch discrimination in fluorescent in situ hybridization using different types of nucleic acids  |h [Elektronische Daten]  |c [Fontenete Silvia, Barros Joana, Madureira Pedro, Figueiredo Céu, Wengel Jesper, Azevedo Filipe] 
520 3 |a 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. 
540 |a Springer-Verlag Berlin Heidelberg, 2015 
690 7 |a FISH  |2 nationallicence 
690 7 |a Oligonucleotides  |2 nationallicence 
690 7 |a Nucleic acids  |2 nationallicence 
690 7 |a Microbiology  |2 nationallicence 
690 7 |a Mismatch discrimination  |2 nationallicence 
700 1 |a Silvia  |D Fontenete  |u LEPABE—Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465, Porto, Portugal  |4 aut 
700 1 |a Joana  |D Barros  |u LEPABE—Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465, Porto, Portugal  |4 aut 
700 1 |a Pedro  |D Madureira  |u ICBAS, Institute of Biomedical Sciences Abel Salazar, University of Porto, 4050-313, Porto, Portugal  |4 aut 
700 1 |a Céu  |D Figueiredo  |u IPATIMUP, Institute of Molecular Pathology and Immunology of the University of Porto, 4200-465, Porto, Portugal  |4 aut 
700 1 |a Jesper  |D Wengel  |u Nucleic Acid Center, Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, 5230, Odense M, Denmark  |4 aut 
700 1 |a Filipe  |D Azevedo  |u LEPABE—Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465, Porto, Portugal  |4 aut 
773 0 |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/9(2015-05-01), 3961-3969  |x 0175-7598  |q 99:9<3961  |1 2015  |2 99  |o 253 
856 4 0 |u https://doi.org/10.1007/s00253-015-6389-4  |q text/html  |z Onlinezugriff via DOI 
898 |a BK010053  |b XK010053  |c XK010000 
900 7 |a Metadata rights reserved  |b Springer special CC-BY-NC licence  |2 nationallicence 
908 |D 1  |a research-article  |2 jats 
949 |B NATIONALLICENCE  |F NATIONALLICENCE  |b NL-springer 
950 |B NATIONALLICENCE  |P 856  |E 40  |u https://doi.org/10.1007/s00253-015-6389-4  |q text/html  |z Onlinezugriff via DOI 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Silvia  |D Fontenete  |u LEPABE—Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465, Porto, Portugal  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Joana  |D Barros  |u LEPABE—Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465, Porto, Portugal  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Pedro  |D Madureira  |u ICBAS, Institute of Biomedical Sciences Abel Salazar, University of Porto, 4050-313, Porto, Portugal  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Céu  |D Figueiredo  |u IPATIMUP, Institute of Molecular Pathology and Immunology of the University of Porto, 4200-465, Porto, Portugal  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Jesper  |D Wengel  |u Nucleic Acid Center, Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, 5230, Odense M, Denmark  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Filipe  |D Azevedo  |u LEPABE—Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465, Porto, Portugal  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/9(2015-05-01), 3961-3969  |x 0175-7598  |q 99:9<3961  |1 2015  |2 99  |o 253