Comparative evaluation of Taqman real-time PCR and semi-nested VP1 PCR for detection of enteroviruses in clinical specimens

J Clin Virol. 2010 Sep;49(1):73-4. doi: 10.1016/j.jcv.2010.06.022. Epub 2010 Jul 27.

Abstract

Background: Molecular diagnostic tests to detect enterovirus in clinical specimens usually target highly conserved sites in the 5'-non-translated region, allowing detection of all members of the genus. The sequences in the 5'-NTR do not correlate with serotype, but PCR and sequencing of the VP1 region can be used for typing; this system has largely replaced traditional antigenic typing.

Objective: To investigate the relative performance of two common enterovirus assays.

Study design: We compared the relative sensitivities of Taqman real-time RT-PCR (rRT-PCR) and a VP1 semi-nested PCR (RT-snPCR) assay in which sequencing the VP1 amplicon also provides typing information.

Results: There was 89% concordance between the two methods among the 371 clinical specimens tested (74 positive in both assays and 257 negative in both assays). Twenty-seven rRT-PCR-negative/VP1-positive specimens were confirmed positive by sequencing; 13 specimens were rRT-PCR-positive/VP1-negative.

Conclusions: The results suggest that either assay can produce satisfactory results, but that using both assays in parallel should provide the highest sensitivity for clinical diagnostic testing.

Publication types

  • Comparative Study
  • Evaluation Study

MeSH terms

  • Capsid Proteins / genetics*
  • Cerebrospinal Fluid / virology
  • Enterovirus / classification
  • Enterovirus / genetics
  • Enterovirus / isolation & purification*
  • Enterovirus Infections / blood*
  • Enterovirus Infections / cerebrospinal fluid*
  • Enterovirus Infections / diagnosis
  • Enterovirus Infections / virology
  • Feces / virology
  • Humans
  • Nasopharynx / virology
  • Polymerase Chain Reaction
  • RNA, Viral / analysis
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity

Substances

  • Capsid Proteins
  • RNA, Viral