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Impact of real-time polymerase chain reaction in the diagnosis of acute meningitis in children

Abstract number: P2209

Tormo N., Latorrre J.C., Garcia T., Clari M.A., Navarro D., Gimeno C.

Introduction: Meningitis is one of the most important infectious syndromes in medical emergencies in hospitals, due to its high morbidity and mortality. Meningitis caused by viruses normally present milder symptoms and are the most common, however bacterial meningitis causes greater mortality. Both aetiologies must be quickly distinguished in order to manage patients properly.

Objectives: To compare real-time polymerase chain reaction (PCR) with traditional cell and bacterial cultures in the diagnosis of acute meningitis.

Materials and Methods: 173 cerebrospinal fluid samples (CSF) and blood culture samples from children aged 55 days to 14 years old, collected from January 2006 to November 2007, were tested in the Microbiology Unit of Clinic Hospital of Valencia (Spain).

All samples were cultured for bacterial and viral detection following the laboratory standard procedure. Chocolate agar, Sabouraud-chloramphenicol agar and Brain Heart Infusion broth were used for bacterial culture; and for viral detection the samples were inoculated in RD, VERO and MRC-5 cell cultures.

Nucleic acid was extracted using a BioRobot EZ1 Workstation with EZ1 Viral Mini Kit v2.0 (Qiagen) and with EZ1 DNA Blood 200 ml Kit (Qiagen) for RNA and DNA respectively.

Multiplex real-time PCR was used to test each sample for Neisseria meningitidis and Streptococcus pneumoniae using Realcycler MENE-01 (Ingenie Molecular) in the Smart Cycler® II System (Cepheid). Enteroviruses were tested using real-time reverse transcription-PCR with the Enterovirus Analyte Specific Reagent kit and the Smart Cycler® II System or Xpert EV kit and a GeneXpert® thermocycler (Cepheid).

Results: Fifty-three patients (30.6%) were positive, 44 of them were positive for enterovirus; 35 cases were positive only by PCR, 4 only by cell culture, and 5 by both methods. Sensitivity was 90.9% for the real-time PCR method but only 20.4% for cell culture.

PCR method was able to detect 6 patients infected with N. meningitidis and 3 with S. pneumoniae, whether by culture only 4 cases for N. meningitidis (only 1 from a CSF sample) and 1 case for S. pneumoniae (by blood culture) were detected. PCR showed 100% sensitivity when compared to CSF culture (11%) and blood culture (44%).

Conclusion: Real-time PCR has shown to be a powerful and rapid diagnostic tool in acute meningitis; bacterial and viral aetiologies were characterised in 4 to 5 hours, helping to reduce antibiotic treatment and hospitalisation time.

Session Details

Date: 19/04/2008
Time: 00:00-00:00
Session name: 18th European Congress of Clinical Microbiology and Infectious Diseases
Subject:
Location: Barcelona, Spain
Presentation type:
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