Application of locked nucleic acid-based probes in fluorescence in situ hybridization

• Type: Article in International Scientific Journal
• Date: 2016

Authors

Sílvia Fontenete (Author) FEUP; Daniel Carvalho (Author) Other; Nuno Guimarães (Author) FEUP; Pedro Madureira (Author) Other; Céu Figueiredo (Author) FMUP; Jesper Wengel (Author) Other; Nuno Filipe Azevedo (Author) FEUP

Journal

Title: Applied Microbiology and Biotechnology

Vol. 100

Pages: 5897-5906

ISSN: 0175-7598

Publisher: Springer Nature

Indexing

ISI Web of Knowledge - 14 Citations

Scopus - 13 Citations

Associated Projects

LEPAE/CEFT - RL2 - (Bio) Chemical Engineering: Multi-Scale Approaches for Sustainable Environment and Health

DNA mimics - Desenvolvimento e aplicação de mímicos de DNA para a rápida identificação de microrganismos patogénicos

Other information

Authenticus ID: P-00K-97R

DOI: 10.1007/s00253-016-7429-4

Abstract (EN): 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.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 10