Amplicor enterovirus polymerase chain reaction in patients with aseptic meningitis: a sensitive test limited by amplification inhibitors

Arch Pathol Lab Med. 1999 Oct;123(10):882-4. doi: 10.5858/1999-123-0882-AEPCRI.

Abstract

Objective: To evaluate the sensitivity and specificity of reverse-transcription polymerase chain reaction (RT-PCR) and routine cell culture for the detection of enterovirus in cerebrospinal fluid.

Methods: Thirty-eight cerebrospinal fluid specimens were included. Cell culture was inoculated immediately and incubated for 14 days. An aliquot was kept frozen for Amplicor RT-PCR. Chart review was performed to determine the validity of the results.

Results: Nine of 38 specimens were positive for enterovirus by culture, and 14 were positive by RT-PCR. There were 7 discrepancies between the 2 methods. Six specimens were positive by RT-PCR and negative by the culture method. The 1 culture-positive but RT-PCR--negative specimen was determined to contain PCR inhibitors. All discrepant results were confirmed as true positives by chart review. Patients whose cerebrospinal fluid was negative by both methods had a final diagnosis other than enterovirus infection.

Conclusion: Amplicor PCR is more sensitive than cell culture (93.3% vs. 60%) and is very specific. With the incorporation of appropriate controls for the detection of amplification inhibitors, RT-PCR could be a valuable tool in the diagnosis of enteroviral meningitis.

MeSH terms

  • Cell Culture Techniques / methods
  • Cell Line, Transformed
  • Cerebrospinal Fluid / virology*
  • Enterovirus / genetics*
  • Enterovirus / isolation & purification
  • Enterovirus Infections / cerebrospinal fluid
  • Enterovirus Infections / diagnosis*
  • Evaluation Studies as Topic
  • Fluorescent Antibody Technique, Indirect
  • Humans
  • Meningitis, Aseptic / cerebrospinal fluid
  • Meningitis, Aseptic / diagnosis*
  • RNA, Viral / analysis*
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Virus Cultivation

Substances

  • RNA, Viral