title: Development of a diagnostic real-time polymerase chain reaction assay for the detection of invasive Haemophilus influenzae in clinical samples.
abstract: Since the introduction of the Haemophilus influenzae serotype b vaccine, invasive H. influenzae disease has become dominated by nontypeable (NT) strains. Several widely used molecular diagnostic methods have been shown to lack sensitivity or specificity in the detection of some of these strains. Novel real-time assays targeting the fucK, licA, and ompP2 genes were developed and evaluated. The fucK assay detected all strains of H. influenzae tested (n = 116) and had an analytical sensitivity of 10 genome copies/polymerase chain reaction (PCR). This assay detected both serotype b and NT H. influenzae in 12 previously positive specimens (culture and/or bexA PCR) and also detected H. influenzae in a further 5 of 883 culture-negative blood and cerebrospinal fluid (CSF) samples. The fucK assay has excellent potential as a diagnostic test for detection of typeable and nontypeable strains of invasive H. influenzae in clinical samples of blood and CSF.
authors.affiliation: Epidemiology and Molecular Biology Unit and Irish Meningococcal and Meningitis Reference Laboratory, The Children's University Hospital, Temple Street, Dublin, Ireland. kenneth.meyler@cuh.ie
authors.forename: Kenneth L
authors.initials: KL
authors.lastname: Meyler
authors.forename: Mary
authors.initials: M
authors.lastname: Meehan
authors.forename: Desiree
authors.initials: D
authors.lastname: Bennett
authors.forename: Robert
authors.initials: R
authors.lastname: Cunney
authors.forename: Mary
authors.initials: M
authors.lastname: Cafferkey
mesh_terms: D006192:Haemophilus Infections / Q000175:diagnosis* / Q000382:microbiology; D006193:Haemophilus influenzae / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity