title: Molecular detection of late-onset neonatal sepsis in premature infants using small blood volumes: proof-of-concept.
abstract: BACKGROUND
  Conventional blood culture is still the gold standard for sepsis diagnosis but results are not immediately available and pathogens are only detected in approximately 25% of cases. New molecular assays for the detection of blood stream pathogens are promising diagnostic tools.
  
  
  OBJECTIVE
  The aim of the study was to adapt and evaluate a multiplex PCR system using 100 µl blood. -
  
  
  METHODS
  46 blood specimens of very low birth weight infants (818 ± 242 g) with suspected sepsis were analyzed using the Roche SeptiFast MGRADE PCR with a modified DNA extraction protocol and software handling tool for decreased blood volume requirements.
  
  
  RESULTS
  In the non-infected group, 5/21 infants had a positive PCR result with coagulase-negative staphylococci. All pathogens detected in the blood culture positive group (n = 15) were also detected by PCR. In addition, 4/6 patients had a positive PCR result in the clinical sepsis group (clinical and laboratory signs of sepsis but negative blood culture). Overall, the PCR was demonstrated to have a higher sensitivity (90.5%; 95%CI 68.2-98.3%) in comparison to blood culture (71.4%; 95%CI 47.7-87.8%) including clinical sepsis cases, even though it had a lower specificity (80.0%; 95%CI 58.7-92.4% versus 100.0%; 95%CI 83.4-100.0%).
  
  
  CONCLUSIONS
  These first data demonstrate the usability and potential benefit of this multiplex PCR using a modified DNA extraction for the rapid detection of nosocomial sepsis in preterm infants in addition to blood culture.
authors.affiliation: Department of Pediatrics and Adolescent Medicine, Medical University of Vienna, Vienna, Austria.
authors.forename: David C
authors.initials: DC
authors.lastname: Kasper
authors.forename: Ipek
authors.initials: I
authors.lastname: Altiok
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Mechtler
authors.forename: Judith
authors.initials: J
authors.lastname: Böhm
authors.forename: Julia
authors.initials: J
authors.lastname: Straub
authors.forename: Michaela
authors.initials: M
authors.lastname: Langgartner
authors.forename: Arnold
authors.initials: A
authors.lastname: Pollak
authors.forename: Kurt R
authors.initials: KR
authors.lastname: Herkner
authors.forename: Angelika
authors.initials: A
authors.lastname: Berger
mesh_terms: D057205:Automation, Laboratory; D001431:Bacteriological Techniques; D015415:Biomarkers / Q000097:blood; D001800:Blood Specimen Collection; D003428:Cross Infection / Q000097:blood / Q000175:diagnosis* / Q000382:microbiology; D004269:DNA, Bacterial / Q000097:blood*; D004271:DNA, Fungal / Q000097:blood*; D005865:Gestational Age; D006801:Humans; D007231:Infant, Newborn; D007234:Infant, Premature / Q000097:blood*; D007235:Infant, Premature, Diseases / Q000097:blood / Q000175:diagnosis* / Q000382:microbiology; D019102:Infant, Very Low Birth Weight / Q000097:blood; D060885:Multiplex Polymerase Chain Reaction; D011237:Predictive Value of Tests; D060888:Real-Time Polymerase Chain Reaction; D018805:Sepsis / Q000097:blood / Q000175:diagnosis* / Q000382:microbiology; D012984:Software
chemical_list: D015415:Biomarkers; D004269:DNA, Bacterial; D004271:DNA, Fungal