New reverse transcription-PCR assay for rapid and sensitive detection of enterovirus genomes in cerebrospinal fluid specimens of patients with aseptic meningitis

Abstract

Enterovirus (EV) detection by a new commercially available reverse transcription (RT)-PCR assay (Penter RT-PCR test) was compared with EV isolation from cell cultures and with EV detection by an in-house RT-PCR assay. Of the 54 cerebrospinal fluid specimens collected during a summer outbreak of aseptic meningitis, 52% were positive by cell culture versus 76% by in-house RT-PCR assay and 80% by the new RT-PCR test (52 versus 76 versus 80%; P = 0.003). This new reliable EV RNA detection test is suitable for clinical diagnosis of EV-related meningitis and may improve the management of EV-related neurological syndromes.

FIG. 1.

Distribution of optical density values obtained by detection of EV RNA in 54 CSF samples from patients with aseptic meningitis. RNA detection was by a new one-step PCR method (Penter RT-PCR) with colorimetric microwell detection assay. The dashed line represents the mean OD value (mean OD ± standard deviation, 0.13 ± 0.05) below which the assay was interpreted as negative (cutoff value). •, OD values obtained from EV culture-negative CSF specimens (n = 26); ○, OD values obtained from CSF samples negative by cell culture and in-house RT-PCR assay (n = 2); ▴, OD values obtained from EV culture-positive CSF samples (n = 28).