Multicenter evaluation of the ENTEROVIRUS R-gene 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.


OBJECTIVE
A new CE-marked real-time RT-PCR assay (ENTEROVIRUS R-gene, Argene) was evaluated in two university hospital virology laboratories.


METHODS
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 microl. 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.
Pillet
Sylvie
S
Laboratoire de Bactériologie-Virologie, GIMAP EA 3064 and University Hospital of Saint-Etienne, 42023 Saint-Etienne Cedex 2, France.
Billaud
Geneviève
G
Omar
Shabir
S
Lina
Bruno
B
Pozzetto
Bruno
B
Schuffenecker
Isabelle
I
D002555:Cerebrospinal Fluid / Q000821:virology; D004770:Enterovirus / Q000235:genetics*; D004769:Enterovirus Infections / Q000175:diagnosis / Q000821:virology*; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D010849:Picornaviridae / Q000235:genetics; D012367:RNA, Viral / Q000302:isolation & purification; D015203:Reproducibility of Results; D012137:Respiratory System / Q000821:virology; D012189:Retrospective Studies; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012703:Serotyping / Q000379:methods*
D012367:RNA, Viral