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

Abstract 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™, 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™ 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  TCID 50 /50 μl. Cross-reactivity was observed with four human rhinoviruses. On 59 CSF specimens analyzed prospectively, the results of the ENTEROVIRUS R-gene™ 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™ 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™ 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.