BACKGROUND: Enteroviruses (EV) and parechoviruses (HPeV) are the most common 
causes of aseptic meningitis, encephalitis and sepsis-like syndrome in neonates. 
Detection by nucleic acid amplification methods improves patient management.
OBJECTIVE: Development of a real-time PCR assay on a LightCycler for 
simultaneous detection of EV, HPeV and an internal control to monitor 
inhibition.
STUDY DESIGN: We investigated the value of the new assay, prospectively, in a 
variety of samples from patients suspected of having viral meningitis or 
sepsis-like syndrome.
RESULTS: The assay detected 64 EV serotypes and HPeV types 1-4. Of 186 patients, 
63 (33.9%) were EV positive and 18 (9.7%) HPeV positive in one or more samples. 
In 43 of 159 feces and 6 of 57 throat samples viral culture and PCR were 
positive. With real-time PCR 27 extra EV and 19 HPeV positives were found. Blood 
and CSF were present from 33 patients. In 19 patients blood and CSF were 
positive, one was only positive in CSF, two were only positive in blood, 11 were 
negative. From 96 patients CSF and/or blood samples were tested and compared to 
results in throat and/or feces samples. Forty patients were EV-PCR and 14 
HPeV-PCR positive in blood and/or CSF. All of these were confirmed by a positive 
PCR for the respective virus in feces and/or throat.
CONCLUSIONS: Simultaneous detection of EV and HPeV with this two-step real-time 
PCR is specific, faster and more sensitive than viral culture. All systemic 
infections (blood or CSF positive) were confirmed in feces. Culture is no longer 
necessary for clinical diagnosis and should only be performed on PCR-positive 
samples to obtain isolates for typing purposes. Application of this assay is an 
important improvement for patient management since the outcome of the analysis 
is available within the time frame of clinical decision-making.