Detection of Enteroviruses in the Cerebrospinal Fluid by Polymerase Chain Reaction: Prospective Study of Impact on the Management of Hospitalized Children

Spicher, Virginie Masserey; Berclaz, Pierre-Yves; Cheseaux, Jean‐Jacques; Morandi, Pierre-Alain; Suter, Susanne; Wunderli, Werner; Siegrist, Claire‐Anne

doi:10.1177/000992280003900402

Cited by 24 publications

(17 citation statements)

References 11 publications

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“…The development of enterovirus RT-PCR might avoid the pitfalls of such assumptions. Furthermore, as observed elsewhere [Romero, 1999, Sawyer, 1999, Spicher et al, 2000, early reports of a positive RT-PCR result could save considerable health resources by avoiding unnecessary antibiotic therapy and reducing the length of hospital stays.…”

Section: Discussionmentioning

confidence: 69%

Circulation of enteroviruses and persistence of meningitis cases in the winter of 1999–2000

Chambon

Archimbaud

Bailly

et al. 2001

Journal of Medical Virology

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The seasonal incidence of enterovirus meningitis was analyzed in a prospective study of patients admitted for suspected meningitis from October 1, 1998 to April 30, 2000. In-house reverse transcription-polymerase chain reaction (RT-PCR) in cerebrospinal fluid (CSF) was used irrespective of cytological results. Fifty-two (45.2%) of the 115 patients had positive RT-PCR in CSF, including 44/86 children (51.2%) and 8/29 adults (27.6%). Six of the 52 (11.5%) had no pleocytosis. The numbers of CSF specimens with a predominance of lymphocytes or a predominance of neutrophils were closely similar. In 33 of the positive patients, an enterovirus, mainly echoviruses type 6 (48%) and 30 (24%), was recovered in one or more specimens. Sixteen cases of enteroviral meningitis were observed between November 1999 and March 2000 as against 2 cases between November 1998 and March 1999, showing that the disease persisted through the winter months of 1999-2000. During the same period, 96 enterovirus isolates were recovered from clinical specimens from other patients. The number of isolates was higher in the winter of 1999-2000 (P< 0.01) than in the winter of 1998-1999, indicating that the risk of enterovirus infection increased significantly in winter 1999-2000. Sixteen patients had aseptic meningitis, made a rapid recovery and had an enterovirus in throat swabs and stools (9/16) or in one of the two (7/16). RT-PCR was not requested. Nine patients were admitted during the cold months. The clinical management of both adult and child patients could be improved by year-round use of enterovirus generic RT-PCR.

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Section: Discussionmentioning

confidence: 69%

Circulation of enteroviruses and persistence of meningitis cases in the winter of 1999–2000

Chambon

Archimbaud

Bailly

et al. 2001

Journal of Medical Virology

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“…Rapid diagnosis is essential for appropriate management since an early positive enterovirus diagnosis can avoid inappropriate treatment (such as antibiotic or antiviral therapy) [Romero, 1999;Sawyer, 1999] and unnecessary ancillary tests, and in addition can reduce the duration of hospitalization [Romero, 1999;Spicher et al, 2000]. Rapid diagnosis is also important because effective antivirals like pleconaril 1 are now becoming available for the treatment of enterovirus infection [Rotbart and Webster, 2001;Archimbaud et al, 2003].…”

Section: Introductionmentioning

confidence: 99%

Improved diagnosis on a daily basis of enterovirus meningitis using a one‐step real‐time RT‐PCR assay

Archimbaud

Mirand

Chambon

et al. 2004

Journal of Medical Virology

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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 TCID50/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 TCID50/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.

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“…The use of commercially available (6,11,12,20) and in-house assay methods (6) is currently common in molecular diagnostic virology laboratories and often results in fewer tests being performed, less medication being used, and earlier release from the hospital than with previous diagnostic methods for patients with CSF samples with positive RT-PCR results for EV (EV RT-PCR) and negative Gram stains (4,8,10,14,15).…”

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confidence: 99%

Correlation of Cerebrospinal Fluid (CSF) Cell Counts and Elevated CSF Protein Levels with Enterovirus Reverse Transcription-PCR Results in Pediatric and Adult Patients

Mulford

Buller²,

Arens³

et al. 2004

J Clin Microbiol

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We found that in the group of patients aged >2 months, the absence of pleocytosis was highly predictive of a negative RT-PCR result. Elevated CSF protein level was not a good predictor of RT-PCR positivity for enterovirus and did not add to the diagnostic sensitivity or specificity of pleocytosis.Enteroviruses (EVs) cause annual seasonal epidemics of aseptic meningitis in temperate climates and are the most common cause of this clinical syndrome in the United States. The infections are most clinically noticeable in children, but they are also clearly present in adults (9). The viruses are maintained within the human population in tropical climates year-round and at low but detectable levels in both children and adults during the cold months in temperate climates (3).Reverse transcription (RT)-PCR has become the method of choice for the rapid and efficient diagnosis of EV infections from cerebrospinal fluid (CSF). The use of commercially available (6,11,12,20) and in-house assay methods (6) is currently common in molecular diagnostic virology laboratories and often results in fewer tests being performed, less medication being used, and earlier release from the hospital than with previous diagnostic methods for patients with CSF samples with positive RT-PCR results for EV (EV RT-PCR) and negative Gram stains (4,8,10,14,15).Whether CSF pleocytosis plays an important role in defining aseptic meningitis or in prioritizing the differential diagnosis has been investigated but is still not completely clear (7,14,17). In this era of rapidly rising health care costs, it is important, if possible, to limit laboratory-based testing by encouraging syndrome-directed testing and actively discouraging testing that is not consistent with the clinical picture or with other laboratory results. Additionally, quality control within the laboratory can be enhanced by monitoring laboratory-based data to look for important correlations that will provide insight for clinicians and laboratory workers with regard to future testing requests.During the 2001, 2002, and 2003 EV seasons, we collected all available CSF cell count data and CSF protein data (available for 2001 and 2002 only) for patients for whom EV RT-PCR of CSF samples had been requested. Our suspicion that CSF with a nucleated cell count of Յ5 per mm 3 from patients more than 2 months old was not likely to be RT-PCR positive proved to be true, although a large number of CSF samples with high cell counts did not test positive for EV. MATERIALS AND METHODSCSF specimens (n ϭ 1,706) were submitted to the St. Louis Children's Hospital Virology Laboratory during the 2001, 2002, and 2003 EV seasons (roughly May to November of each year) for EV RT-PCR testing. Relevant CSF cell count data was available from the laboratory information system for 728 of the patients who were seen at Washington University Medical Center and tested by EV RT-PCR. Of the patients included in the study, 549 were pediatric (Յ18 years of age) and 179 were adults. The data analyzed included patient age, EV RT-PC...

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Detection of Enteroviruses in the Cerebrospinal Fluid by Polymerase Chain Reaction: Prospective Study of Impact on the Management of Hospitalized Children

Cited by 24 publications

References 11 publications

Circulation of enteroviruses and persistence of meningitis cases in the winter of 1999–2000

Circulation of enteroviruses and persistence of meningitis cases in the winter of 1999–2000

Improved diagnosis on a daily basis of enterovirus meningitis using a one‐step real‐time RT‐PCR assay

Correlation of Cerebrospinal Fluid (CSF) Cell Counts and Elevated CSF Protein Levels with Enterovirus Reverse Transcription-PCR Results in Pediatric and Adult Patients

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