Comparative genome analysis identifies novel nucleic acid diagnostic targets for use in the specific detection of Haemophilus influenzae

Helena Coughlan; Kate Reddington; Nina Tuite; Teck Wee Boo; Martin Cormican; Louise Barrett; Terry J Smith; Eoin Clancy; Thomas Barry

doi:10.1016/j.diagmicrobio.2015.06.013

Comparative genome analysis identifies novel nucleic acid diagnostic targets for use in the specific detection of Haemophilus influenzae

Diagn Microbiol Infect Dis. 2015 Oct;83(2):112-6. doi: 10.1016/j.diagmicrobio.2015.06.013. Epub 2015 Jun 23.

Authors

Helena Coughlan¹, Kate Reddington², Nina Tuite², Teck Wee Boo³, Martin Cormican⁴, Louise Barrett⁵, Terry J Smith⁶, Eoin Clancy⁶, Thomas Barry²

Affiliations

¹ Nucleic Acid Diagnostics Research Laboratory (NADRL), Microbiology, School of Natural Sciences, National University of Ireland, Galway, Ireland; Biomedical Diagnostics Institute Programme, National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland.
² Nucleic Acid Diagnostics Research Laboratory (NADRL), Microbiology, School of Natural Sciences, National University of Ireland, Galway, Ireland.
³ Department of Clinical Microbiology, University College Hospital, Galway, Ireland.
⁴ School of Medicine, National University of Ireland, Galway, Ireland.
⁵ National Centre of Sensor Research, School of Physical Sciences, Biomedical Diagnostics Institute, Dublin City University, Glasnevin, Dublin 9, Ireland.
⁶ Biomedical Diagnostics Institute Programme, National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland; Molecular Diagnostics Research Group (MDRG), School of Natural Sciences, National University of Ireland, Galway, Ireland.

PMID: 26166209
DOI: 10.1016/j.diagmicrobio.2015.06.013

Abstract

Haemophilus influenzae is recognised as an important human pathogen associated with invasive infections, including bloodstream infection and meningitis. Currently used molecular-based diagnostic assays lack specificity in correctly detecting and identifying H. influenzae. As such, there is a need to develop novel diagnostic assays for the specific identification of H. influenzae. Whole genome comparative analysis was performed to identify putative diagnostic targets, which are unique in nucleotide sequence to H. influenzae. From this analysis, we identified 2H. influenzae putative diagnostic targets, phoB and pstA, for use in real-time PCR diagnostic assays. Real-time PCR diagnostic assays using these targets were designed and optimised to specifically detect and identify all 55H. influenzae strains tested. These novel rapid assays can be applied to the specific detection and identification of H. influenzae for use in epidemiological studies and could also enable improved monitoring of invasive disease caused by these bacteria.

Keywords: Genome comparison; H. influenzae; Real-time PCR; phoB; pstA.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

ATP-Binding Cassette Transporters / genetics
Bacterial Proteins / genetics
Bacteriological Techniques / methods*
Computational Biology*
DNA, Bacterial / genetics*
Genome, Bacterial*
Haemophilus Infections / diagnosis*
Haemophilus Infections / microbiology
Haemophilus influenzae / genetics
Haemophilus influenzae / isolation & purification*
Humans
Molecular Epidemiology / methods
Real-Time Polymerase Chain Reaction / methods*
Time Factors

Substances

ATP-Binding Cassette Transporters
Bacterial Proteins
DNA, Bacterial
phosphate-specific transport protein A, bacteria
PhoB protein, Bacteria