708. Introduction of a Novel Parechovirus RT-PCR in a Clinical Virology Laboratory
Session: Abstracts: Virology
Friday, October 22, 2010
Background: Parechovirus (PEV) is an important cause of meningitis and sepsis-like illness in children. We designed and introduced PEV RT-PCR in routine testing of cerebrospinal fluid (CSF) and blood specimens submitted to our Clinical Virology Laboratory.

Methods: RT-PCR was designed to detect all known PEV within the 5’ untranslated region. Test validation included PEV RT-PCR testing of enterovirus (ENT) and rhinovirus (RHN) culture harvests as well as retrospective clinical samples from July 2007 to December 2008 (n=867). Since May 2009, this assay has been performed in real time on samples submitted for ENT RT-PCR. Following IRB approval, medical records from patients who had PEV detected were reviewed. 

Results: 142 viral cultures previously identified as ENT by immunofluorescence were tested by PEV and ENT RT-PCR; 3 were strongly positive for PEV and 139 for ENT.  Four samples positive for ENT were also weakly positive for PEV suggesting low cross-reactivity. Among 107 viral cultures identified as RHN by cytopathic effect and growth characteristics, 2 were positive by PEV RT-PCR and negative by RHN RT-PCR.  Of the 867 retrospective clinical samples, 6 were positive for PEV and 71 for ENT without cross-reactivity. The PEV RT-PCR has been performed prospectively since May 2009 in parallel with ENT RT-PCR. Of 499 clinical specimens (409 CSF, 90 blood), 48 specimens from 46 patients were positive for ENT (9.6% positivity) and 17 specimens from 15 patients for PEV (3.4% positivity). The addition of PEV to our standard ENT RT-PCR test request improved pathogen recovery by 35%. During this study, PEV represented 26% of samples with either ENT or PEV detected.  Twelve PEV positive pediatric patients hospitalized at our medical center had medical records reviewed. Average age was 29 days (7-57 days); average duration of fever was 3.4 days (1-5 days).  Two of 11 patients had proteinorachia >120mg/L; one had CSF pleocytosis. One of the 12 patients had severe encephalitis but no patient died.

Conclusion: PEV RT-PCR should be considered part of routine testing of CSF samples from young children and infants.


Subject Category: V. Virology including clinical and basic studies of viral infections, including hepatitis

Speakers:
Christian Renaud, MD, MSc , Infectious Disease Sciences, Fred Hutchinson Cancer Research, Seattle, WA
Elle Ficken , Infectious Diseases, Seattle Children's Hospital, Seattle, WA
Anne Cent , Laboratory Medecine, Seattle Children's Hospital, Seattle, WA
Lawrence Corey, MD , University of Washington, Fred Hutchinson Cancer Research Center, Seattle, WA
Janet A. Englund, MD, FIDSA , Seattle Children's Research Institute, Seattle, WA
Jane Kuypers, PhD , Laboratory Medicine, University of Washington, Seattle, WA

Disclosures:

C. Renaud, None

E. Ficken, None

A. Cent, None

L. Corey, None

J. A. Englund, None

J. Kuypers, None

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