title: HIRA-TAN: a real-time PCR-based system for the rapid identification of causative agents in pneumonia.
abstract: Identification of the causative pathogen(s) of pneumonia would allow the selection of effective antibiotics and thus reduce the mortality rate and the emergence of drug-resistant pathogens. To identify such pathogens and to obtain these benefits, it is necessary that a clinical test is rapid, accurate, easily performed, and cost-effective. Here, we devised a PCR-based test, named HIRA-TAN, which is able to discriminate therapeutic targets from commensal organisms (e.g. Streptococcus pneumoniae or Haemophilus influenzae) and to detect foreign organisms (e.g. Mycoplasma pneumoniae or Legionella pneumophila) in the sputum. The utility of this system was validated in a prospective study, using sputum samples from patients with pneumonia. 568 patients were enrolled and the HIRA-TAN assay identified the causative pathogens with an accuracy of 96.7% for H. influenzae; 93.2% for Pseudomonas aeruginosa; 80.6% for Klebsiella pneumoniae; 90.9% for Moraxella catarrhalis; 87.5% for Escherichia coli; 78.1% for MRSA and 91.6% for S. pneumoniae. Overall the HIRA-TAN procedure was able to identify the causative pathogens of pneumonia in 60% of the cases. Additionally, this procedure was able to determine when the pneumonia-causing organism was a commensal organism or a foreign organism in a single assay. The HIRA-TAN approach yielded reproducible results and provided valuable information to plan the course of treatment of pneumonia. Through the rapid identification of the causative pathogens, the HIRA-TAN will promote targeted treatments for pneumonias.
  
  
  BACKGROUND
  UMIN000001694.
authors.affiliation: Department of Respiratory Medicine, Saitama Medical University, Moroyama-machi, Saitama 350-0495, Japan. Electronic address: mycobacteriumtuberculosis@hotmail.com.
authors.forename: Takashi
authors.initials: T
authors.lastname: Hirama
authors.affiliation: Department of Respiratory Medicine, Saitama Medical University, Moroyama-machi, Saitama 350-0495, Japan.
authors.forename: Shohei
authors.initials: S
authors.lastname: Minezaki
authors.affiliation: Department of Respiratory Medicine, Saitama Medical University, Moroyama-machi, Saitama 350-0495, Japan.
authors.forename: Takefumi
authors.initials: T
authors.lastname: Yamaguchi
authors.affiliation: Central Laboratory, Saitama Medical University, 38 Morohongo, Moroyama-machi, Saitama 350-0495, Japan.
authors.forename: Etsuko
authors.initials: E
authors.lastname: Kishi
authors.affiliation: Department of Respiratory Medicine, Saitama Medical University, Moroyama-machi, Saitama 350-0495, Japan; Department of General Internal Medicine, Saitama Medical University, Moroyama-machi, Saitama 350-0495, Japan.
authors.forename: Keiji
authors.initials: K
authors.lastname: Kodama
authors.affiliation: Department of Respiratory Medicine, Saitama Medical University, Moroyama-machi, Saitama 350-0495, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Egashira
authors.affiliation: Department of Respiratory Medicine, Saitama Medical University International Medical Center, Hidaka, Saitama 350-1298, Japan.
authors.forename: Kunihiko
authors.initials: K
authors.lastname: Kobayashi
authors.affiliation: Department of Respiratory Medicine, Saitama Medical University, Moroyama-machi, Saitama 350-0495, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Nagata
authors.affiliation: Tsurugashima Ikenodai Hospital, Tsurugashima, Saitama 350-2200, Japan.
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Ishii
authors.affiliation: Department of Emergency and Acute Medicine, Saitama Medical University International Medical Center, Hidaka, Saitama 350-1298, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Nemoto
authors.affiliation: Kan-etsu Hospital, Tsurugashima, Saitama 350-2213, Japan.
authors.forename: Masahiko
authors.initials: M
authors.lastname: Tanaka
authors.affiliation: Pulmonary Division, Department of Medicine, Keio University School of Medicine, Shinjuku, Tokyo 160-8582, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Fukunaga
authors.affiliation: Department of Respiratory Medicine, Saitama Medical University, Moroyama-machi, Saitama 350-0495, Japan.
authors.forename: Minoru
authors.initials: M
authors.lastname: Kanazawa
authors.affiliation: Department of Respiratory Medicine, Saitama Medical University, Moroyama-machi, Saitama 350-0495, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Hagiwara
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001419:Bacteria / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D018410:Pneumonia, Bacterial / Q000175:diagnosis*; D011446:Prospective Studies; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D013183:Sputum / Q000382:microbiology; D013559:Symbiosis; D014216:Triacetoneamine-N-Oxyl; D055815:Young Adult
chemical_list: D014216:Triacetoneamine-N-Oxyl
keywords: Commensal organism; Foreign organism; HIRA-TAN; Pneumonia; Real-time PCR