title: Detection of Neisseria meningitidis DNA from skin lesion biopsy using real-time PCR: usefulness in the aetiological diagnosis of purpura fulminans.
abstract: OBJECTIVE
  The present study evaluated the usefulness of a real-time polymerase chain reaction (rtPCR) assay for the detection of Neisseria meningitidis (Nm) and genogrouping on skin lesion biopsies in patients with purpura fulminans (PF).
  
  
  METHODS
  Retrospective single-centre study.
  
  
  METHODS
  Adult and paediatric intensive care units at the University Hospital of Rouen.
  
  
  METHODS
  All patients admitted between January 2000 and January 2006, with a final diagnosis of PF and for which a skin biopsy and blood cultures were performed, were included.
  
  
  METHODS
  Skin biopsy and blood cultures were used for culture and rtPCR.
  
  
  RESULTS
  Thirty-four patients fulfilled the criteria (27 children and 7 adults). Nm rtPCR performed on skin biopsy was positive in 100% (34/34) of cases, compared with only 14.7% (5/34) for skin culture (p=0.0001). rtPCR genogrouping on skin biopsy was positive in 58.8% (20/34) of the cases compared with 14.7% (5/34) for skin culture (p=0.0013). For patients (n=17) in whom rtPCR was performed both on blood and skin biopsy, skin biopsy gave a significantly higher rate of Nm detection [100% (17/17) vs. 58.8% (10/17); p=0.023] and genogroup characterisation [76.5% (13/17) vs. 35.3% (6/17); p=0.045] than blood. We encountered no specimen with culture-positive and rtPCR-negative results (negative predictive value of rtPCR 100%).
  
  
  CONCLUSIONS
  In suspected PF cases, skin biopsy is more reliable to identify Nm and its genogroup than blood or, probably, CSF, especially when PCR methods are used. This could help the implementation of public health interventions, especially concerning a vaccination policy.
authors.affiliation: Service de Pédiatrie Néonatale et Réanimation, Centre Hospitalier Universitaire de Rouen, 1, Rue de Germont, 76031, Rouen Cedex, France. pierre.staquet@chu-rouen.fr.
authors.forename: Pierre
authors.initials: P
authors.lastname: Staquet
authors.affiliation: Groupe de Recherche sur les Antimicrobiens et les Microorganismes (G.R.A.M. EA 2656, I.F.R. 23), Université de Rouen, U.F.R. Médecine-Pharmacie, Rouen, France.|Service de Bactériologie, University Hospital of Rouen, Rouen, France.
authors.forename: Ludovic
authors.initials: L
authors.lastname: Lemee
authors.affiliation: Service de Dermatologie, University Hospital of Rouen, Rouen, France.
authors.forename: Eva
authors.initials: E
authors.lastname: Verdier
authors.affiliation: Service de Réanimation Médicale, University Hospital of Rouen, Rouen, France.
authors.forename: Guy
authors.initials: G
authors.lastname: Bonmarchand
authors.affiliation: Service de Pédiatrie Néonatale et Réanimation, Centre Hospitalier Universitaire de Rouen, 1, Rue de Germont, 76031, Rouen Cedex, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Laudenbach
authors.affiliation: Service de Pédiatrie Néonatale et Réanimation, Centre Hospitalier Universitaire de Rouen, 1, Rue de Germont, 76031, Rouen Cedex, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Michel
authors.affiliation: Groupe de Recherche sur les Antimicrobiens et les Microorganismes (G.R.A.M. EA 2656, I.F.R. 23), Université de Rouen, U.F.R. Médecine-Pharmacie, Rouen, France.|Service de Bactériologie, University Hospital of Rouen, Rouen, France.
authors.forename: Jean-Francois
authors.initials: JF
authors.lastname: Lemeland
authors.affiliation: Service de Pédiatrie Néonatale et Réanimation, Centre Hospitalier Universitaire de Rouen, 1, Rue de Germont, 76031, Rouen Cedex, France.
authors.forename: Stephane
authors.initials: S
authors.lastname: Marret
authors.affiliation: Service de Pédiatrie Néonatale et Réanimation, Centre Hospitalier Universitaire de Rouen, 1, Rue de Germont, 76031, Rouen Cedex, France.
authors.forename: Thierry
authors.initials: T
authors.lastname: Blanc
mesh_terms: D000328:Adult; D001706:Biopsy; D001769:Blood / Q000382:microbiology; D002648:Child; D004269:DNA, Bacterial / Q000302:isolation & purification*; D006801:Humans; D011695:IgA Vasculitis / Q000382:microbiology*; D007362:Intensive Care Units; D015278:Intensive Care Units, Pediatric; D009345:Neisseria meningitidis / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012189:Retrospective Studies; D012867:Skin / Q000737:chemistry / Q000382:microbiology*
chemical_list: D004269:DNA, Bacterial