Evaluation of a rapid real-time RT-PCR assay for detection of enterovirus RNA in cerebrospinal fluid specimens

J Clin Virol. 2002 Jul:25 Suppl 1:S39-43. doi: 10.1016/s1386-6532(02)00032-x.

Abstract

Background: We previously described the characteristics of a single-tube real-time enterovirus reverse transcriptase polymerase chain reaction (RT-PCR) assay based on a fluorogenic probe and primers directed to highly conserved sequences in the 5'-untranslated region (UTR) of the enterovirus genome.

Objectives: To evaluate the performance of the assay on a larger number of cerebrospinal fluid (CSF) specimens from patients suspected of having viral meningitis.

Study design: Real-time enterovirus RT-PCR and viral culture were performed on CSF specimens received from March 2000 to November 2001. Patient records were retrospectively reviewed for final clinical diagnosis.

Results: From the 186 CSF specimens tested, culture was positive for enterovirus in 31 cases, whereas real-time RT-PCR detected enterovirus RNA in 45 CSF specimens. The sensitivity of real-time RT-PCR in relation to the clinical diagnosis of viral meningitis was 72.6%, whereas the sensitivity of viral culture reached only 57.4%. Enterovirus RNA was also found in a number of specimens with low leukocyte counts.

Conclusions: We confirm that the real-time enterovirus RT-PCR assay for CSF specimens is significantly more sensitive than viral culture.

Publication types

  • Comparative Study

MeSH terms

  • Cerebrospinal Fluid / virology
  • Enterovirus / genetics
  • Enterovirus / growth & development
  • Enterovirus / isolation & purification*
  • Enterovirus Infections / cerebrospinal fluid
  • Enterovirus Infections / diagnosis*
  • Enterovirus Infections / virology
  • Evaluation Studies as Topic
  • Humans
  • Leukocyte Count
  • Meningitis, Viral / cerebrospinal fluid
  • Meningitis, Viral / diagnosis*
  • Meningitis, Viral / virology
  • Polymerase Chain Reaction / methods*
  • RNA, Viral / analysis*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sensitivity and Specificity

Substances

  • RNA, Viral