Korean J Lab Med.  2006 Feb;26(1):9-13. 10.3343/kjlm.2006.26.1.9.

Detection of Enterovirus in Cerebrospinal Fluid by Real-Time Nested Reverse Transcription Polymerase Chain Reaction

• Heo SR
• Jin SK
• Chang HE
• Park KU
• Song J
• Kim EC

• KMID: 2143190
• DOI: http://doi.org/10.3343/kjlm.2006.26.1.9

Abstract

BACKGROUND: Enterovirus is a common cause of aseptic meningitis, respiratory disease and nonspecific febrile illness. The conventional methods for laboratory diagnosis of enterovirus infections have been virus culture and serotyping by an immunofluorecent test. We studied a new and more rapid approach for enterovirus detection in cerebrospinal fluid (CSF) by real-time nested PCR.
METHODS
This study was performed on 50 CSF specimens from patients suspected of aseptic meningitis. Enterovirus was detected in CSF by PCRs for 3 different targets and real-time nested PCR. Enterovirus culture was also performed in 44 CSF specimens.
RESULTS
The positive rate of PCRs for each of the 3 different targets was 26.0%, 40.0%, or 46.0%, and that of real-time nested PCR was 86.0%. Only 6.8% were positive in culture. Thus, the positive rate of real-time nested PCR was much higher than other methods.
CONCLUSIONS
Our study revealed that the real-time nested PCR should be useful for diagnosis of enterovirus infections because of a high sensitivity and rapid detection.

Keyword

Enterovirus; Real-time polymerase chain reaction; Nested PCR; Reverse Transcription PCR; Aseptic meningitis

MeSH Terms

Cerebrospinal Fluid*
Clinical Laboratory Techniques
Diagnosis
Enterovirus Infections
Enterovirus*
Humans
Meningitis, Aseptic
Polymerase Chain Reaction*
Real-Time Polymerase Chain Reaction
Reverse Transcription*
Serotyping

Figure

• 

  Fig. 1. Detection of enterovirus by RT-PCR methods for 3 different targets using CSF specimens. (A) The amplified products of first method using EV1-F and EV1-R primers are 114 bp. (B) The amplified products of second method using EV2-F and EV2-R primers are 154 bp. (C) The amplified products of third method using EV3-F and EV3-R primers are 149 bp. Lane 1, negative control; lane 2, positive control; lane M, 100 bp DNA size marker; lane 3–12, specimens.

Reference

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