The DOI matches its Crossref record; 26 of the 26 references it cites were found in a registry or an index.
Improved diagnosis on a daily basis of enterovirus meningitis using a one‐step real‐time RT‐PCR assay
Abstract
The detection of the enterovirus genome in cerebrospinal fluid (CSF) by PCR techniques has proved to be more sensitive than traditional cell culture for the diagnosis of enterovirus meningitis. However, PCR assays are time consuming and labor intensive, particularly if separate hybridization steps are used to confirm the specificity of positive findings. The aim of this study was to develop a one‐step real‐time RT‐PCR assay with LightCycler (LC) technology that was sensitive, rapid, and easy to perform in routine practice. The enterovirus detection limit was determined by testing 10‐fold limiting dilution series of cell culture stocks with the echovirus 25 (E‐25) prototype strain and with the third European Union Quality Control Concerted Action (EU‐QCCA) enterovirus proficiency panel. A total of 100 CSF specimens were investigated in a comparative study. With the E‐25 strain, the detection limit of the real‐time assay was 286 TCID 50/ml (50% tissue culture infective dose). When samples of the EU‐QCCA panel were tested, our assay gave identical results (detection limit down to 3.6 TCID 50/ml) to those of the reference laboratory, which used one‐step RT‐PCR assay. When CSF specimens were tested, there was a correlation between the real‐time assay and the conventional in‐house assay in 96 of 100 CSFs tested. This one‐step real‐time assay allows rapid enterovirus detection in CSF since results are obtained in 3 hr as against 36 hr with the “in‐house” RT‐PCR assay. This new assay is now being used in routine practice, and allows diagnosis on a daily basis. J. Med. Virol. 74:604–611, 2004. © 2004 Wiley‐Liss, Inc.
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“The reason that only Enteroviridae were detected in this study and that no etiologic agents were identified in two-thirds of cases is likely that cultures on CSF are not sensitive for other viruses [13].”
From Paediatric Investigators Collaborative Network on Infections in Canada (PICNIC) study of aseptic meningitis (2006) · cites this paper as [13]
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Metadata
- Authors
- Archimbaud, C., Mirand, A., Chambon, M., Regagnon, C., Bailly, J., Peigue‐Lafeuille, H., & Henquell, C.
- Affiliations
- Centre Hospitalier Universitaire de Clermont-Ferrand (FR)
- Corresponding author
- Archimbaud, Christine
- Venue
- Journal of Medical Virology
- Publisher
- Wiley
- Published
- 2004-10-13
- Volume
- 74
- Issue
- 4
- Pages
- 604-611
- ISSN
- 0146-6615 (ISSN-L)
- 1096-9071
- OpenAlex
- W2125092867
- Cited by
- 28
- References
- 26
- Citations by year
- 2024: 1 · 2023: 1 · 2021: 1 · 2018: 1 · 2017: 1 · 2013: 5 · 2012: 1
- FWCI
- 2.86
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Medical Virology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Infectious Diseases › Viral Infections and Immunology Research
- Health Sciences › Medicine › Infectious Diseases › Infectious Encephalopathies and Encephalitis
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Molecular Biology Techniques and Applications
- Keywords
- enterovirus meningitis, enterovirus detection, cerebrospinal fluid, LightCycler, detection limit, rapid diagnosis
- MeSH terms
- Cerebrospinal Fluid, Enterovirus Infections, Enterovirus, Humans, Meningitis, Viral, RNA, Viral, Sensitivity and Specificity, Time Factors, Polymerase Chain Reaction
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