Hybridization-based detection of Helicobacter pylori at human body temperature using advanced Locked Nucleic Acid (LNA) probes

• Type: Article in International Scientific Journal
• Date: 2013

Authors

Sílvia Fontenete (Author) FEUP; Nuno Guimarães (Author) FEUP; Marina Leite (Author) Other; Céu Figueiredo (Author) Other; Jesper Wengel (Author) Other; Nuno Filipe Azevedo (Author) FEUP

Journal

Title: PLoS ONE

Vol. 8 No. 11

Pages: 1-11

ISSN: 1932-6203

Publisher: Public Library of Science

Indexing

ISI Web of Knowledge - 35 Citations

Scopus - 34 Citations

Associated Projects

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

Other information

Authenticus ID: P-006-JHZ

DOI: 10.1371/journal.pone.0081230

Abstract (EN): The understanding of the human microbiome and its influence upon human life has long been a subject of study. Hence, methods that allow the direct detection and visualization of microorganisms and microbial consortia (e. g. biofilms) within the human body would be invaluable. In here, we assessed the possibility of developing a variant of fluorescence in situ hybridization (FISH), named fluorescence in vivo hybridization (FIVH), for the detection of Helicobacter pylori. Using oligonucleotide variations comprising locked nucleic acids (LNA) and 2'-O-methyl RNAs (2'OMe) with two types of backbone linkages (phosphate or phosphorothioate), we were able to successfully identify two probes that hybridize at 37 degrees C with high specificity and sensitivity for H. pylori, both in pure cultures and in gastric biopsies. Furthermore, the use of this type of probes implied that toxic compounds typically used in FISH were either found to be unnecessary or could be replaced by a non-toxic substitute. We show here for the first time that the use of advanced LNA probes in FIVH conditions provides an accurate, simple and fast method for H. pylori detection and location, which could be used in the future for potential in vivo applications either for this microorganism or for others.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 11