DIAGNOSIS OF VENTRICULAR DRAINAGE‐RELATED BACTERIAL MENINGITIS BY BROAD‐RANGE REAL‐TIME POLYMERASE CHAIN REACTION

OBJECTIVE To compare a broad-range real-time polymerase chain reaction (PCR) diagnostic strategy with culture to evaluate additional effects on the etiological diagnosis and the quantification of the bacterial load during the course of ventricular drainage-related bacterial meningitis (VR-BM). METHODS We applied a PCR that targeted conserved regions of the 16S ribosomal ribonucleic acid gene to cerebrospinal fluid (CSF) samples from patients with external ventricular drainage or a ventriculoperitoneal shunt during the course of VR-BM. We compared the PCR results with CSF cultures. A total of 350 routine CSF samples were consecutively collected from 86 patients. The CSF deoxyribonucleic acid was automatically purified and subjected to PCR. Amplicons from the PCR samples that were positive for VR-BM were subsequently deoxyribonucleic acid sequenced for final identification. Clinical data were extracted from patient files. RESULTS Sixteen patients had at least one VR-BM-positive sample as diagnosed from culture or PCR. Nineteen episodes were diagnosed with signs of VR-BM (n = 16 patients) or were determined to be contaminated (n = 3 patients). Four episodes of VR-BM were diagnosed via PCR alone and were predominantly caused by gram-negative pathogens, five episodes were diagnosed via culture alone, and seven episodes were diagnosed via both culture and PCR. Five patients had mixed infections. Overall, 71 samples were positive for VR-BM as indicated by either one or both of the methods. Eighteen CSF samples were VR-BM positive as indicated by culture alone, and 21 CSF samples were positive as indicated via PCR alone. CONCLUSIONS Culture supplemented with broad-range, real-time PCR may increase the number of etiologically diagnosed VR-BM episodes, particularly when these are caused by gram-negative bacteria.

[1]  M. Quigley,et al.  Biofilm-related infections of cerebrospinal fluid shunts. , 2006, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[2]  J. Møller,et al.  Broad-range real time PCR and DNA sequencing for the diagnosis of bacterial meningitis , 2006, Scandinavian journal of infectious diseases.

[3]  R. Tubbs,et al.  Polymerase Chain Reaction for the Rapid Detection of Cerebrospinal Fluid Shunt or Ventriculostomy Infections , 2005, Neurosurgery.

[4]  E. Kuijper,et al.  Bacterial meningitis caused by the use of ventricular or lumbar cerebrospinal fluid catheters. , 2005, Journal of neurosurgery.

[5]  S. Clarke,et al.  Interlaboratory Comparison of PCR-Based Identification and Genogrouping of Neisseria meningitidis , 2005, Journal of Clinical Microbiology.

[6]  P. Hsueh,et al.  Direct Detection of Bacterial Pathogens in Brain Abscesses by Polymerase Chain Reaction Amplification and Sequencing of Partial 16S Ribosomal Deoxyribonucleic Acid Fragments , 2004, Neurosurgery.

[7]  T. Czech,et al.  Early diagnosis of external ventricular drainage infection: results of a prospective study , 2003, Journal of neurology, neurosurgery, and psychiatry.

[8]  A. Fox,et al.  Employment of broad-range 16S rRNA PCR to detect aetiological agents of infection from clinical specimens in patients with acute meningitis--rapid separation of 16S rRNA PCR amplicons without the need for cloning. , 2003, Journal of applied microbiology.

[9]  W. Poon,et al.  Failure of regular external ventricular drain exchange to reduce cerebrospinal fluid infection: result of a randomised controlled trial , 2002, Journal of neurology, neurosurgery, and psychiatry.

[10]  E. Connolly,et al.  Ventriculostomy-related Infections: A Critical Review of the Literature , 2002, Neurosurgery.

[11]  J. Moore,et al.  Risk Assessment Models and Contamination Management: Implications for Broad-Range Ribosomal DNA PCR as a Diagnostic Tool in Medical Bacteriology , 2002, Journal of Clinical Microbiology.

[12]  R. Proctor,et al.  Coagulase-negative staphylococci , 2001 .

[13]  Y. Chuang,et al.  Community-Acquired Acinetobacter Meningitis in Adults , 2000, Infection.

[14]  Didier Raoult,et al.  16S Ribosomal DNA Sequence Analysis of a Large Collection of Environmental and Clinical Unidentifiable Bacterial Isolates , 2000, Journal of Clinical Microbiology.

[15]  R. Borrow,et al.  Contamination and Sensitivity Issues with a Real-Time Universal 16S rRNA PCR , 2000, Journal of Clinical Microbiology.

[16]  D. Low,et al.  Nosocomial bacterial meningitis, including central nervous system shunt infections. , 1999, Infectious disease clinics of North America.

[17]  S. Nikkari,et al.  Diagnosis of Meningococcal Meningitis by Broad-Range Bacterial PCR with Cerebrospinal Fluid , 1998, Journal of Clinical Microbiology.

[18]  M. Nguyen,et al.  Antibiotic-resistant Acinetobacter meningitis in neurosurgical patients. , 1994, Neurosurgery.

[19]  T. Bannerman,et al.  Update on clinical significance of coagulase-negative staphylococci , 1994, Clinical Microbiology Reviews.

[20]  D. Ross,et al.  Differentiation of aseptic and bacterial meningitis in postoperative neurosurgical patients. , 1988, Journal of neurosurgery.

[21]  H. Abdel-salam Direct PCR assay for detection ofNeisseria meningitidis in human cerebrospinal fluid , 2008, Folia Microbiologica.

[22]  L. Saravolatz,et al.  Broad-range bacterial polymerase chain reaction for early detection of bacterial meningitis. , 2003, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[23]  Neil Hunter,et al.  Determination of bacterial load by real-time PCR using a broad-range (universal) probe and primers set. , 2002, Microbiology.

[24]  Thomas D. Schmittgen,et al.  Real-Time Quantitative PCR , 2002 .

[25]  James R. Cole,et al.  rrndb: the Ribosomal RNA Operon Copy Number Database , 2001, Nucleic Acids Res..

[26]  Y. Chuang,et al.  Mixed Infection in Adult Bacterial Meningitis , 2000, Infection.

[27]  W. Poon,et al.  CSF antibiotic prophylaxis for neurosurgical patients with ventriculostomy: a randomised study. , 1998, Acta neurochirurgica. Supplement.