Development and application of a novel peptide nucleic acid (PNA) probe for the specific detection of Helicobacter pylori

Guimaraes, N.M., Azevedo, N.F., Figueiredo, C., Keevil, C.W. and Vieira, M.J. (2006) Development and application of a novel peptide nucleic acid (PNA) probe for the specific detection of Helicobacter pylori. Helicobacter, 11 (Abs.10.10), 383-383. (doi:10.1111/j.1523-5378.2006.00419_12.x).

Abstract

The standard methods for accurate diagnosis of Helicobacter pylori infection consist in either culturing of the pathogen and/or concordant positive results obtained by histology and the rapid urease test or the 13C-urea breath test (UBT), as none of these diagnostic tests except bacterial culturing are 100% specific, fluorescence in situ hybridization (FISH) might be a good complement. In this work, the development of a new peptide nucleic acid (PNA) FISH probe for the detection of H. pylori is reported. The 15-mer PNA probe was connected to an Alexa Fluor 546 dye and target a specific 16S rRNA sequence of the bacterium. PNA probes are a recent technology that has been shown to provide for additional specificity and sensitivity in FISH procedures when compared to the standard DNA counterparts. The probe was tested against several H. pylori and non-H. pylori strains, and was shown to be specific for the microorganism of interest. This technique was optimized for different types of supports such as slides, membrane filters, and also coupons of various materials where H. pylori was present. Tests performed showed a better sensitivity of the probe than the standard plating procedures for H. pylori detection. We are currently optimizing the application of this new probe for paraffin-embedded gastric biopsy specimens. When completely optimized, this technique could be a useful method, as it is rapid, sensitive, and specific, for an even more accurate diagnostic of H. pylori infection, and could also be used for distinguishing H. pylori from other Helicobacter species.

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Contributors

Author: C.W. Keevil

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