title: Comparative genome analysis identifies novel nucleic acid diagnostic targets for use in the specific detection of Haemophilus influenzae.
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.
authors.affiliation: 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.
authors.forename: Helena
authors.initials: H
authors.lastname: Coughlan
authors.affiliation: Nucleic Acid Diagnostics Research Laboratory (NADRL), Microbiology, School of Natural Sciences, National University of Ireland, Galway, Ireland.
authors.forename: Kate
authors.initials: K
authors.lastname: Reddington
authors.affiliation: Nucleic Acid Diagnostics Research Laboratory (NADRL), Microbiology, School of Natural Sciences, National University of Ireland, Galway, Ireland.
authors.forename: Nina
authors.initials: N
authors.lastname: Tuite
authors.affiliation: Department of Clinical Microbiology, University College Hospital, Galway, Ireland.
authors.forename: Teck Wee
authors.initials: TW
authors.lastname: Boo
authors.affiliation: School of Medicine, National University of Ireland, Galway, Ireland.
authors.forename: Martin
authors.initials: M
authors.lastname: Cormican
authors.affiliation: National Centre of Sensor Research, School of Physical Sciences, Biomedical Diagnostics Institute, Dublin City University, Glasnevin, Dublin 9, Ireland.
authors.forename: Louise
authors.initials: L
authors.lastname: Barrett
authors.affiliation: 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.
authors.forename: Terry J
authors.initials: TJ
authors.lastname: Smith
authors.affiliation: 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.
authors.forename: Eoin
authors.initials: E
authors.lastname: Clancy
authors.affiliation: Nucleic Acid Diagnostics Research Laboratory (NADRL), Microbiology, School of Natural Sciences, National University of Ireland, Galway, Ireland.
authors.forename: Thomas
authors.initials: T
authors.lastname: Barry
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D001431:Bacteriological Techniques / Q000379:methods*; D019295:Computational Biology; D004269:DNA, Bacterial / Q000235:genetics*; D016680:Genome, Bacterial; D006192:Haemophilus Infections / Q000175:diagnosis* / Q000382:microbiology; D006193:Haemophilus influenzae / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D017720:Molecular Epidemiology / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D013997:Time Factors
chemical_list: D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D004269:DNA, Bacterial; C078408:phosphate-specific transport protein A, bacteria; C061278:PhoB protein, Bacteria
keywords: Genome comparison; H. influenzae; Real-time PCR; phoB; pstA