Evaluation of a rapid real-time RT-PCR assay for detection of enterovirus RNA in cerebrospinal fluid specimens

Verstrepen, Walter A.; Bruynseels, Peggy; Mertens, An

doi:10.1016/s1386-6532(02)00032-x

Cited by 95 publications

(71 citation statements)

References 15 publications

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“…Detection of EV is usually performed by isolation of the virus in cell culture, reverse transcription PCR (RT-PCR), or real-time RT-PCR ( 11 , 14 – 16 ). Currently, RT-PCR is used routinely worldwide to diagnose EV infection because of its sensitivity, specificity, and ability to detect highly conserved 5′ noncoding regions of the human EV genome ( 15 , 17 , 18 ). For determining subtype, the neutralization test is the standard diagnostic tool and is generally reliable, but it is also labor-intensive, time-consuming, and may fail to identify an isolate ( 16 , 19 ).…”

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confidence: 99%

Accuracy of Diagnostic Methods and Surveillance Sensitivity for Human Enterovirus, South Korea, 1999–2011

Hyeon¹,

Hwang²,

Kim³

et al. 2013

Emerg. Infect. Dis.

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The epidemiology of enteroviral infection in South Korea during 1999–2011 chronicles nationwide outbreaks and changing detection and subtyping methods used over the 13-year period. Of 14,657 patients whose samples were tested, 4,762 (32.5%) samples were positive for human enterovirus (human EV); as diagnostic methods improved, the rate of positive results increased. A seasonal trend of outbreaks was documented. Genotypes enterovirus 71, echovirus 30, coxsackievirus B5, enterovirus 6, and coxsackievirus B2 were the most common genotypes identified. Accurate test results correlated clinical syndromes to enterovirus genotypes: aseptic meningitis to echovirus 30, enterovirus 6, and coxsackievirus B5; hand, foot and mouth disease to coxsackievirus A16; and hand, foot and mouth disease with neurologic complications to enterovirus 71. There are currently no treatments specific to human EV infections; surveillance of enterovirus infections such as this study provides may assist with evaluating the need to research and develop treatments for infections caused by virulent human EV genotypes.

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confidence: 99%

Accuracy of Diagnostic Methods and Surveillance Sensitivity for Human Enterovirus, South Korea, 1999–2011

Hyeon¹,

Hwang²,

Kim³

et al. 2013

Emerg. Infect. Dis.

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“…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.…”

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confidence: 99%

GeneXpert Enterovirus Assay: One-Year Experience in a Routine Laboratory Setting and Evaluation on Three Proficiency Panels

et al. 2008

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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) ...

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“…Moreover, failure of detection of an EV serotype can sometimes be attributed to the presence of a small amount of infectious virus in CSF specimens [5,45]. The use of molecular techniques described in this report improved the diagnosis of EV meningitis in our laboratory [22,58]. Previous studies have shown that the PCR techniques including nested PCR are less time consuming and more sensitive than viral culture based techniques in the diagnosis of enteroviral meningitis [27,52].…”

Section: Discussionmentioning

confidence: 92%

Enteroviral Central Nervous System Infections in Children of the Region of Monastir, Tunisia: Diagnosis, Laboratory Findings of Cerebrospinal Fluid and Clinical Manifestations

et al. 2012

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Human enteroviruses (HEV) are one of the major causes of central nervous system (CNS) infections in pediatrics. A prospective study was conducted to assess the epidemiological, clinical, and laboratory characteristics of enterovirus (EV) infections of the CNS in children under 15-years-old, suspected of having viral CNS infections and admitted to the Pediatric Department of Monastir University Hospital, Tunisia. Enteroviral RNA was detected by 5 0 NCR nested RT-PCR assay in 33 % (20 out of 60) of cerebrospinal fluid specimens, whereas only six samples (10 %) were EV positive in cell culture. EV-positive patients were clustered according to their clinical manifestations, predominantly diagnosed as aseptic meningitis (65 %) and meningoencephalitis (20 %). Fever, headache, vomiting, and neck stiffness were the most pronounced symptoms. Pleocytosis with the predominance of lymphocytes was observed in 60 % of EV positive specimens. Although patients suffering from EV infections were encountered throughout the year, most occurred during spring and summer months. Using VP1-2A nested RT-PCR and sequence analysis, three of the 20 positive HEV were identified as Echovirus (E)-9. This is the first report of a cluster of aseptic meningitis cases caused by E-9 in Monastir.

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Evaluation of a rapid real-time RT-PCR assay for detection of enterovirus RNA in cerebrospinal fluid specimens

Cited by 95 publications

References 15 publications

Accuracy of Diagnostic Methods and Surveillance Sensitivity for Human Enterovirus, South Korea, 1999–2011

Accuracy of Diagnostic Methods and Surveillance Sensitivity for Human Enterovirus, South Korea, 1999–2011

GeneXpert Enterovirus Assay: One-Year Experience in a Routine Laboratory Setting and Evaluation on Three Proficiency Panels

Enteroviral Central Nervous System Infections in Children of the Region of Monastir, Tunisia: Diagnosis, Laboratory Findings of Cerebrospinal Fluid and Clinical Manifestations

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