Comparison of an in-house real-time RT-PCR assay with a commercial assay for detection of enterovirus RNA in clinical samples

Eur J Clin Microbiol Infect Dis. 2012 May;31(5):715-9. doi: 10.1007/s10096-011-1364-1. Epub 2011 Jul 31.

Abstract

Molecular detection of enterovirus (EV) RNA based on PCR methods is a quicker and more sensitive approach than culture methods. At present, different PCR-based methods for EV RNA detection are available, but comparisons of results obtained according to the different approaches are limited. We evaluated an in-house real-time RT-PCR assay with a commercialized TaqMan real-time RT-PCR kit for detection of EV. Consecutive clinical specimens from paediatric patients less than 6 years old with clinical suspicion of EV infection were analyzed between July and November 2010. After RNA extraction, samples were amplified both by the real-time RT-PCR commercial assay and the in-house assay. A total of 19 of 132 patients (14.4%) involving 20 samples (14 plasma samples and 6 CSF) were positive in at least one of the two assays. The sensitivity of the in-house assay when the MutaPLATE® assay was used as a reference was 90% (IC 95%; 74.35-100) and the specificity was 100% (IC 95%; 99.63-100). Cts results of two methods were statistically correlated (r = 0.774; P = 0.01). In conclusion, these two real-time RT-PCR assays are rapid and easy methods for detection of EV.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cerebrospinal Fluid / virology
  • Child
  • Child, Preschool
  • Clinical Laboratory Techniques / methods*
  • Enterovirus / genetics
  • Enterovirus / isolation & purification*
  • Enterovirus Infections / diagnosis*
  • Female
  • Humans
  • Infant
  • Infant, Newborn
  • Male
  • Molecular Diagnostic Techniques / methods*
  • Plasma / virology
  • RNA, Viral / genetics
  • RNA, Viral / isolation & purification*
  • Real-Time Polymerase Chain Reaction / methods*
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Virology / methods

Substances

  • RNA, Viral