Multicenter evaluation of the ENTEROVIRUS R-gene (TM) real-time RT-PCR assay for the detection of enteroviruses in clinical specimens

Background: The rapid molecular diagnosis of enteroviral meningitis has been shown important for an adequate management of the patients. Objectives: A new CE-marked real-time RT-PCR assay (ENTEROVIRUS R-gene (TM), Argene) was evaluated in two university hospital virology laboratories. Study design: Reactivity, analytical sensitivity and specificity were evaluated using 54 prototype and 173 clinical human enterovirus (HEV) strains, a 12-sample HEV proficiency panel, and 30 non-HEV microorganisms. The clinical performance of the ENTEROVIRUS R-gene (TM) assay was evaluated by testing 197 cerebrospinal fluid (CSF) and 103 respiratory specimens, comparatively to the routinely used diagnostic techniques. Results: Sixty-four out of the 65 HEV serotypes tested were detected. The analytical sensitivity ranged between 10(-2.64) and 10(2.39) TCID50/50 mu l. Cross-reactivity was observed with four human rhinoviruses. On 59 CSF specimens analyzed prospectively, the results of the ENTEROVIRUS R-gene (TM) assay showed a 94.8% concordance with those of the Smart enterovirus (EV) assay (Cepheid). On 138 CSF specimens tested retrospectively, the results of the ENTEROVIRUS R-gene (TM) assay showed a 97.1% concordance with those of either the GeneXpert EV assay (Cepheid) or the in-house RT-PCR HEV assays used at the time of specimen collection. On 103 respiratory specimens, the concordance between the results of the ENTEROVIRUS R-gene (TM) assay and those of the routine RT-PCRs or viral culture was 90.2% and 96.1% before and after retest, respectively. Conclusions: The new test was found able to detect a large panel of enterovirus serotypes; it was sensitive when used on clinical specimens; and, easy and rapid to perform on a routine basis. (C) 2009 Elsevier B.V. All rights reserved.