Rapid detection of human parechoviruses in clinical samples by real-time PCR.
BACKGROUND
Human parechoviruses (HPeVs) have been associated with severe conditions such as neonatal sepsis and meningitis in young children. Rapid identification of an infectious agent in such serious conditions in these patients is essential for adequate decision making regarding treatment and hospital stay.


OBJECTIVE
We have developed an HPeV specific real-time PCR assay based on the conserved 5'untranslated region.


METHODS
To determine the detection limit of the assay, serial dilutions of HPeV in vitro RNA were tested in a background of HPeV and EV RNA-negative cerebrospinal fluid (CSF). The specificity was tested by analyzing culture isolates of HPeV 1-6, enterovirus (EV) types, human rhinoviruses (HRVs) and hepatitis A virus (HAV). To establish diagnostic relevance, 522 CSF samples from children <5 years were tested.


RESULTS
The detection limit of the assay was 75 copies of HPeV cDNA per reaction. The assay was highly specific for HPeV, detecting all HPeV types. We identified HPeV infections in CSF of 20 children (3.8%), all with severe conditions such as sepsis and meningitis.


CONCLUSIONS
These results suggest that HPeV screening of paediatric clinical samples should be included in viral diagnostics in suspected cases of neonatal sepsis and meningitis.
Benschop
Kimberley
K
Department of Medical Microbiology, Laboratory of Clinical Virology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands. k.s.benschop@amc.uva.nl
Molenkamp
Richard
R
van der Ham
Alwin
A
Wolthers
Katja
K
Beld
Marcel
M
D020121:5' Untranslated Regions / Q000235:genetics; D002675:Child, Preschool; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008587:Meningitis, Viral / Q000175:diagnosis / Q000821:virology; D030061:Parechovirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D010850:Picornaviridae Infections / Q000175:diagnosis / Q000821:virology; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000134:cerebrospinal fluid* / Q000235:genetics; D018805:Sepsis / Q000175:diagnosis / Q000821:virology; D013045:Species Specificity; D013997:Time Factors
D020121:5' Untranslated Regions; D012367:RNA, Viral