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Rapid detection of human parechoviruses in clinical samples by real-time PCR
Kimberley Benschop1, Richard Molenkamp, Alwin van der Ham
1Department of Medical Microbiology, Laboratory of Clinical Virology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands. k.s.benschop@amc.uva.nl
Insights
A new real-time PCR assay accurately detects human parechoviruses (HPeVs) in children. This rapid identification is crucial for diagnosing severe conditions like neonatal sepsis and meningitis, guiding treatment decisions.
Area of Science:
- Virology
- Molecular Diagnostics
- Pediatric Infectious Diseases
Background:
- Human parechoviruses (HPeVs) are linked to severe neonatal sepsis and meningitis in children.
- Timely identification of infectious agents is critical for effective treatment and patient management.
Purpose of the Study:
- To develop a specific real-time PCR assay for human parechovirus (HPeV) detection.
- To establish a rapid diagnostic tool for HPeV in pediatric clinical samples.
Main Methods:
- A real-time PCR assay was designed targeting the conserved 5' untranslated region of HPeV.
- Assay sensitivity was determined using serial dilutions of HPeV RNA in cerebrospinal fluid (CSF).
- Specificity was evaluated against various viral pathogens, and 522 pediatric CSF samples were analyzed.
Main Results:
- The assay demonstrated a detection limit of 75 copies of HPeV cDNA per reaction.
- High specificity for all tested HPeV types was confirmed.
- HPeV infection was identified in 3.8% of pediatric CSF samples, primarily associated with sepsis and meningitis.
Conclusions:
- HPeV screening should be integrated into routine viral diagnostics for suspected neonatal sepsis and meningitis.
- The developed assay offers a valuable tool for rapid HPeV detection in pediatric populations.
- Early diagnosis of HPeV can significantly impact clinical decision-making and patient outcomes.
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.
Objectives:
We have developed an HPeV specific real-time PCR assay based on the conserved 5'untranslated region.
Study Design:
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.
