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

Christine Archimbaud1, Audrey Mirand, Martine Chambon

  • 1Laboratoire de Virologie Médicale du Centre Hospitalier Universitaire, Faculté de Médecine, Clermont-Ferrand Cedex 1, France. carchimbaud@chu-clermontferrand.fr

Journal of Medical Virology
|October 16, 2004
PubMed

Insights

A new one-step real-time RT-PCR assay offers rapid and sensitive detection of enterovirus meningitis in cerebrospinal fluid (CSF). This method significantly reduces diagnostic time compared to traditional PCR, aiding daily clinical practice.

Area of Science:

  • Virology
  • Molecular Diagnostics
  • Infectious Diseases

Background:

  • Enterovirus meningitis diagnosis traditionally relies on PCR, which is sensitive but time-consuming.
  • Conventional PCR requires separate hybridization steps, increasing labor and time for specificity confirmation.
  • Existing methods pose challenges for rapid, routine diagnosis in clinical settings.

Purpose of the Study:

  • To develop a sensitive, rapid, and user-friendly one-step real-time RT-PCR assay for enterovirus detection.
  • To implement LightCycler (LC) technology for enhanced efficiency in enterovirus diagnostics.
  • To facilitate timely diagnosis of enterovirus meningitis in clinical practice.

Main Methods:

  • Developed a one-step real-time RT-PCR assay utilizing LightCycler technology.
  • Determined assay sensitivity using serial dilutions of echovirus 25 (E-25) and an enterovirus proficiency panel.
  • Compared the real-time assay performance against a conventional in-house RT-PCR assay using 100 CSF specimens.

Main Results:

  • The real-time assay detected as low as 286 TCID50/ml of E-25 and 3.6 TCID50/ml with the EU-QCCA panel.
  • Achieved results consistent with a reference laboratory's one-step RT-PCR assay.
  • Demonstrated a 96% correlation with the conventional assay in 100 CSF samples.
  • Reduced turnaround time to 3 hours from 36 hours compared to the in-house assay.

Conclusions:

  • The developed one-step real-time RT-PCR assay is highly sensitive and rapid for enterovirus detection in CSF.
  • This assay simplifies the diagnostic process, making it suitable for routine clinical use.
  • Enables daily diagnosis of enterovirus meningitis, improving patient management.

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