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

Abstract

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. To evaluate the performance of the assay on a larger number of cerebrospinal fluid (CSF) specimens from patients suspected of having viral meningitis. 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. 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. We confirm that the real-time enterovirus RT-PCR assay for CSF specimens is significantly more sensitive than viral culture.