title: Evaluation of pathogen detection from clinical samples by real-time polymerase chain reaction using a sepsis pathogen DNA detection kit.
abstract: BACKGROUND
  Sepsis is a serious medical condition that requires rapidly administered, appropriate antibiotic treatment. Conventional methods take three or more days for final pathogen identification and antimicrobial susceptibility testing. We organized a prospective observational multicenter study in three study sites to evaluate the diagnostic accuracy and potential clinical utility of the SeptiFast system, a multiplex pathogen detection system used in the clinical setting to support early diagnosis of bloodstream infections.
  
  
  METHODS
  A total of 212 patients, suspected of having systemic inflammatory response syndrome (SIRS) caused by bacterial or fungal infection, were enrolled in the study. From these patients, 407 blood samples were taken and blood culture analysis was performed to identify pathogens. Whole blood was also collected for DNA Detection Kit analysis immediately after its collection for blood culture. The results of the DNA Detection Kit, blood culture and other culture tests were compared. The chosen antimicrobial treatment in patients whose samples tested positive in the DNA Detection Kit and/or blood culture analysis was examined to evaluate the effect of concomitant antibiotic exposure on the results of these analyses.
  
  
  RESULTS
  SeptiFast analysis gave a positive result for 55 samples, while 43 samples were positive in blood culture analysis. The DNA Detection Kit identified a pathogen in 11.3% (45/400) of the samples, compared to 8.0% (32/400) by blood culture analysis. Twenty-three pathogens were detected by SeptiFast only; conversely, this system missed five episodes of clinically significant bacteremia (Methicillin-resistant Staphylococcus aureus (MRSA), 2; Pseudomonas aeruginosa, 1; Klebsiella spp, 1; Enterococcus faecium, 1). The number of samples that tested positive was significantly increased by combining the result of the blood culture analysis with those of the DNA Detection Kit analysis (P = 0.01). Among antibiotic pre-treated patients (prevalence, 72%), SeptiFast analysis detected more bacteria/fungi, and was less influenced by antibiotic exposure, compared with blood culture analysis (P = 0.02).
  
  
  CONCLUSIONS
  This rapid multiplex pathogen detection system complemented traditional culture-based methods and offered some added diagnostic value for the timely detection of causative pathogens, particularly in antibiotic pre-treated patients. Adequately designed intervention studies are needed to prove its clinical effectiveness in improving appropriate antibiotic selection and patient outcomes.
authors: Department of Laboratory Medicine, Nagasaki University School of Medicine, 1-7-1 Sakamoto, Nagasaki City, Nagasaki 852-8501, Japan. kyana-ngs@umin.ac.jp
authors: Katsunori
authors: K
authors: Yanagihara
authors: Yuko
authors: Y
authors: Kitagawa
authors: Masao
authors: M
authors: Tomonaga
authors: Kunihiro
authors: K
authors: Tsukasaki
authors: Shigeru
authors: S
authors: Kohno
authors: Masafumi
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authors: Seki
authors: Hisashi
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authors: Sugimoto
authors: Takeshi
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authors: Shimazu
authors: Osamu
authors: O
authors: Tasaki
authors: Asako
authors: A
authors: Matsushima
authors: Yasuo
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authors: Ikeda
authors: Shinichiro
authors: S
authors: Okamoto
authors: Naoki
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authors: Aikawa
authors: Shingo
authors: S
authors: Hori
authors: Hideaki
authors: H
authors: Obara
authors: Akitoshi
authors: A
authors: Ishizaka
authors: Naoki
authors: N
authors: Hasegawa
authors: Junzo
authors: J
authors: Takeda
authors: Shimeru
authors: S
authors: Kamihira
authors: Kazuyuki
authors: K
authors: Sugahara
authors: Seishi
authors: S
authors: Asari
authors: Mitsuru
authors: M
authors: Murata
authors: Yoshio
authors: Y
authors: Kobayashi
authors: Hiroyuki
authors: H
authors: Ginba
authors: Yoshinobu
authors: Y
authors: Sumiyama
authors: Masaki
authors: M
authors: Kitajima
mesh_terms: D000900:Anti-Bacterial Agents / Q000627:therapeutic use; D001424:Bacterial Infections / Q000097:blood / Q000175:diagnosis / Q000188:drug therapy; D004269:DNA, Bacterial / Q000097:blood; D004271:DNA, Fungal / Q000097:blood; D006801:Humans; D055624:Methicillin-Resistant Staphylococcus aureus / Q000235:genetics; D009181:Mycoses / Q000097:blood / Q000175:diagnosis / Q000188:drug therapy; D011933:Reagent Kits, Diagnostic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018805:Sepsis / Q000097:blood / Q000175:diagnosis* / Q000382:microbiology; D013203:Staphylococcal Infections / Q000097:blood / Q000175:diagnosis; D018746:Systemic Inflammatory Response Syndrome / Q000097:blood / Q000175:diagnosis / Q000382:microbiology
chemical_list: D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D004271:DNA, Fungal; D011933:Reagent Kits, Diagnostic