caa a22 4500 60549990X CHVBK 20210128100553.0 cr unu---uuuuu 210128e20150501xx s 000 0 eng 10.1007/s00253-015-6389-4 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6389-4 Mismatch discrimination in fluorescent in situ hybridization using different types of nucleic acids [Elektronische Daten] [Fontenete Silvia, Barros Joana, Madureira Pedro, Figueiredo Céu, Wengel Jesper, Azevedo Filipe] 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. Springer-Verlag Berlin Heidelberg, 2015 FISH nationallicence Oligonucleotides nationallicence Nucleic acids nationallicence Microbiology nationallicence Mismatch discrimination nationallicence Silvia Fontenete LEPABE—Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465, Porto, Portugal aut Joana Barros LEPABE—Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465, Porto, Portugal aut Pedro Madureira ICBAS, Institute of Biomedical Sciences Abel Salazar, University of Porto, 4050-313, Porto, Portugal aut Céu Figueiredo IPATIMUP, Institute of Molecular Pathology and Immunology of the University of Porto, 4200-465, Porto, Portugal aut Jesper Wengel Nucleic Acid Center, Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, 5230, Odense M, Denmark aut Filipe Azevedo LEPABE—Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465, Porto, Portugal aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/9(2015-05-01), 3961-3969 0175-7598 99:9<3961 2015 99 253 https://doi.org/10.1007/s00253-015-6389-4 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6389-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Silvia Fontenete LEPABE—Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465, Porto, Portugal aut NATIONALLICENCE 700 1- Joana Barros LEPABE—Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465, Porto, Portugal aut NATIONALLICENCE 700 1- Pedro Madureira ICBAS, Institute of Biomedical Sciences Abel Salazar, University of Porto, 4050-313, Porto, Portugal aut NATIONALLICENCE 700 1- Céu Figueiredo IPATIMUP, Institute of Molecular Pathology and Immunology of the University of Porto, 4200-465, Porto, Portugal aut NATIONALLICENCE 700 1- Jesper Wengel Nucleic Acid Center, Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, 5230, Odense M, Denmark aut NATIONALLICENCE 700 1- Filipe Azevedo LEPABE—Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465, Porto, Portugal aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/9(2015-05-01), 3961-3969 0175-7598 99:9<3961 2015 99 253