title: Quantitative fucK gene polymerase chain reaction on sputum and nasopharyngeal secretions to detect Haemophilus influenzae pneumonia.
abstract: A quantitative polymerase chain reaction (PCR) for the fucK gene was developed for specific detection of Haemophilus influenzae. The method was tested on sputum and nasopharyngeal aspirate (NPA) from 78 patients with community-acquired pneumonia (CAP). With a reference standard of sputum culture and/or serology against the patient's own nasopharyngeal isolate, H. influenzae etiology was detected in 20 patients. Compared with the reference standard, fucK PCR (using the detection limit 10(5) DNA copies/mL) on sputum and NPA showed a sensitivity of 95.0% (19/20) in both cases, and specificities of 87.9% (51/58) and 89.5% (52/58), respectively. In a receiver operating characteristic curve analysis, sputum fucK PCR was found to be significantly superior to sputum P6 PCR for detection of H. influenzae CAP. NPA fucK PCR was positive in 3 of 54 adult controls without respiratory symptoms. In conclusion, quantitative fucK real-time PCR provides a sensitive and specific identification of H. influenzae in respiratory secretions.
authors.affiliation: Section of Clinical Bacteriology, Department of Medical Sciences, Uppsala University, S-75185, Uppsala, Sweden.
authors.forename: Guma M K
authors.initials: GM
authors.lastname: Abdeldaim
authors.forename: Kristoffer
authors.initials: K
authors.lastname: Strålin
authors.forename: Per
authors.initials: P
authors.lastname: Olcén
authors.forename: Jonas
authors.initials: J
authors.lastname: Blomberg
authors.forename: Paula
authors.initials: P
authors.lastname: Mölling
authors.forename: Björn
authors.initials: B
authors.lastname: Herrmann
mesh_terms: D000368:Aged; D017714:Community-Acquired Infections / Q000175:diagnosis / Q000382:microbiology; D004269:DNA, Bacterial / Q000302:isolation & purification; D005798:Genes, Bacterial; D006193:Haemophilus influenzae / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D009305:Nasopharynx / Q000382:microbiology*; D018410:Pneumonia, Bacterial / Q000175:diagnosis* / Q000382:microbiology; D011446:Prospective Studies; D012336:RNA, Ribosomal, 16S / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013183:Sputum / Q000382:microbiology*
chemical_list: D004269:DNA, Bacterial; D007074:Immunoglobulin G; D012336:RNA, Ribosomal, 16S