A prospective unblinded comparative evaluation of three assays for the detection of enteroviral RNA performed on 83 positive and 79 negative cerebrospinal fluid samples showed initial and resolved sensitivities of 90.4% and 98.8%, respectively, for the Cepheid GeneXpert enterovirus assay; 94.0% and 97.6%, respectively, for the Argene enterovirus consensus kit; and 100% and 100%, respectively, for an in-house real-time PCR. The initial and resolved specificities were 100% for all three assays.Enteroviruses comprise a large group of immunologically distinct serotypes of viruses belonging to the family Picornaviridae (18). Infection with enteroviruses is associated with protean clinical manifestations, ranging from asymptomatic or mild febrile illness to severe and potentially fatal syndromes, including paralysis, aseptic meningitis, encephalitis, myocarditis, and neonatal systemic infection (1). Enteroviruses are the most common cause of aseptic meningitis in both children and adults and may cause up to 90% of cases of aseptic meningitis for which an etiology is identified (1,14,18,19). The rapid detection and the rapid characterization of enteroviral meningitis are essential for making decisions for patient management and treatment (5,15,17,20). The provision of enterovirus test results on a daily basis can have a substantial impact on health care and has been shown to be highly cost-effective (9,12,17). Since the conventional means of diagnosis of enteroviral meningitis by cell culture from cerebrospinal fluid (CSF) is timeconsuming and expensive and lacks sensitivity, some commercial and several in-house PCR protocols as well as nucleic acid sequence-based amplification assays have been developed during the last 15 years in order to improve the ability to detect enterovirus from CSF (2-4, 6, 7, 11, 13, 16, 20-22, 24-26). The most widely used PCR-based assays for the routine diagnosis of enteroviral meningitis are those that detect PCR products in microtiter wells (6,21,22) and those that use the real-time PCR technology (2,3,13,(24)(25)(26). However, all of these approaches require specially trained laboratory staff, thus limiting the ability of the "real-time" technology to deliver patient results that would be most useful for making patient management decisions STAT.The latest development in the field of the molecular diagnosis of enteroviral meningitis is a fully automated real-time multiplex, reverse transcription-PCR assay, the GeneXpert enterovirus assay (GXEA; Cepheid, Sunnyvale, CA) (10). GXEA is, at present, the only FDA-approved assay for the qualitative detection of enterovirus RNA in CSF. It combines automated nucleic acid sample preparation, amplification, and real-time detection of enteroviral RNA in a disposable, macro/microfluidic cartridge using the GeneXpert Dx system instrument. To date, only one evaluation of GXEA has been published in peer-reviewed journals. A multicenter beta trial with 102 CSF samples obtained from patients with suspected meningitis (34 of whom were enterovirus positive) ...