Detection of enterovirus RNA in cerebrospinal fluid: Comparison of two molecular assays

Crom, Stephanie C. M. de; Obihara, C C; Loon, Anton M. van; Argilagos-Alvarez, A.A.; Peeters, Marcel; Furth, A. Marceline van; Rossen, John W. A.

doi:10.1016/j.jviromet.2011.10.007

Cited by 23 publications

(13 citation statements)

References 31 publications

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“…The purpose of performing our assay in separate reactions is to achieve specific and simultaneous detection, without the risk of cross-contamination. Using this method, 100% sensitivity was obtained for the 3 viruses, and we observed no cross-reactions with any of the viruses analyzed; additionally, specificities with high values were achieved (EV: 96%, HSV1: 100%, and HSV2: 94%), similar to those reported with the end-point and other real-time techniques, and even higher than the specificity achieved with other commercial kits [7,11,[16][17][18][19][20][21] . These results indicate that the sequences used for the primers and probes are specific; in the case of EV, the primers allow the identification of all viruses grouped in the genus, as the target sequence is highly conserved among its members; in the case of HSV1 and HSV2, as the primers are aimed at 2 different targets, this protocol can achieve differential diagnosis between these 2 viruses, without observing the cross-reactions that have been reported in other studies that use different areas of the same target [22][23][24] .…”

supporting

confidence: 83%

Differential Detection of Enterovirus and Herpes Simplex Virus in Cerebrospinal Fluid by Real-Time RT-PCR

et al. 2017

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Background: Enterovirus (EV) and herpes simplex virus 1 and 2 (HSV1 and HSV2) are the main etiologic agents of central nervous system infections. Early laboratory confirmation of these infections is performed by viral culture of the cerebrospinal fluid (CSF), or the detection of specific antibodies in serum (e.g., HSV). The sensitivity of viral culture ranges from 65 to 75%, with a recovery time varying from 3 to 10 days. Serological tests are faster and easy to carry out, but they exhibit cross-reactivity between HSV1 and HSV2. Although molecular techniques are more sensitive (sensitivity >95%), they are more expensive and highly susceptible to cross-contamination. Methods: A real-time RT-PCR for the detection of EV, HSV1, and HSV2 was compared with end-point nested PCR. Results: We tested 87 CSF samples of patients with a clinical diagnosis of viral meningitis or encephalitis. Fourteen samples were found to be positive by RT-PCR, but only 8 were positive by end-point PCR. The RT-PCR showed a specificity range of 94-100%, the negative predictive value was 100%, and the positive predictive value was 62, 100, and 28% for HSV1, HSV2, and EV, respectively. Conclusion: Real-time RT-PCR detected EV, HSV1, and HSV2 with a higher sensitivity and specificity than end-point nested RT-PCR.

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confidence: 83%

Differential Detection of Enterovirus and Herpes Simplex Virus in Cerebrospinal Fluid by Real-Time RT-PCR

et al. 2017

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“…The increase in EV reporting in recent years closely reflects the increase in PCR testing, which was also associated with a lower proportion of samples being submitted to the national reference department for confirmation and typing. The high sensitivity of PCR as compared with cell culture will also have contributed to the number of laboratory-confirmed cases [12][13][14][15], and this trend is likely to continue as more sensitive multiplex assays for simultaneous detection of multiple pathogens in biological samples are adopted by local hospitals. In addition to being cheap, rapid, and sensitive, PCR testing for EV can directly impact on individual patient care by reducing unnecessary antibiotic usage and shortening hospital stays.…”

Section: Discussionmentioning

confidence: 99%

“…PCR testing is cheaper, rapid, requires small amounts of biological material, can be used for multiple pathogens, and can detect almost all EV types, because the viruses share common genetic sequences. For EVs, PCR has greater sensitivity and specificity than viral cultures [12][13][14][15]. Data from these studies show that it is possible to deliver a result within 5-6 h of sample testing, which could, in turn, deliver significant cost savings by reducing unnecessary antimicrobial use and shortening hospital stays.…”

Section: Introductionmentioning

confidence: 99%

Enterovirus infections in England and Wales, 2000–2011: the impact of increased molecular diagnostics

Kadambari

Bukasa

Okike

et al. 2014

Clinical Microbiology and Infection

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There have recently been significant changes in diagnostic practices for detecting enterovirus (EV) infections across England and Wales. Reports of laboratory-confirmed EV infections submitted by National Health Service (NHS) hospital laboratories to Public Health England (PHE) over a 12-year period (2000-2011) were analysed. Additionally, the PHE Virus Reference Department (VRD) electronic database containing molecular typing data from 2004 onwards was interrogated. Of the 13,901 reports, there was a decline from a peak of 2254 in 2001 to 589 in 2006, and then an increase year-on-year to 1634 in 2011. This increase coincided with increasing PCR-based laboratory diagnosis, which accounted for 36% of reported cases in 2000 and 92% in 2011. The estimated annual incidence in 2011 was 3.9/100,000 overall and 238/100,000 in those aged <3 months, who accounted for almost one-quarter of reported cases (n = 2993, 23%). During 2004-2011, 2770 strains were submitted for molecular typing to the VRD, who found no evidence for a predominance of any particular strain. Thus, the recent increase in reported cases closely reflects the increase in PCR testing by NHS hospitals, but is associated with a lower proportion of samples being submitted for molecular typing. The high EV rate in young infants merits further investigation to inform evidence-based management guidance.

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“…25 Detection frequencies for hPeV in diverse cohorts range from 2.3% to 4.6% in cerebrospinal fluid (CSF). [29][30][31] Our aim was to assess prevalence, epidemiology and clinical presentation of CNS infections with hPeV and compare it to that of EV in our patient population. 27,28 Polymerase chain reaction (PCR) has repeatedly been demonstrated to be superior to viral culture in detecting hPeV in clinical specimens.…”

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confidence: 99%

Human Parechovirus Central Nervous System Infections in Southern California Children

Felsenstein

Yang²,

Eubanks³

et al. 2014

Pediatric Infectious Disease Journal

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HPeV infections of the central nervous system occurred mainly in young infants and were more commonly associated with neurologic symptoms at presentation, despite the fact that CSF findings were within the normal range in the vast majority of these cases. HPeV should be included in the differential diagnosis of young children with central nervous system symptoms and sepsis-like illness, even in the presence of normal CSF parameters.

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Detection of enterovirus RNA in cerebrospinal fluid: Comparison of two molecular assays

Cited by 23 publications

References 31 publications

Differential Detection of Enterovirus and Herpes Simplex Virus in Cerebrospinal Fluid by Real-Time RT-PCR

Differential Detection of Enterovirus and Herpes Simplex Virus in Cerebrospinal Fluid by Real-Time RT-PCR

Enterovirus infections in England and Wales, 2000–2011: the impact of increased molecular diagnostics

Human Parechovirus Central Nervous System Infections in Southern California Children

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