Evaluation of a new real-time reverse transcription polymerase chain reaction assay for detection of norovirus in fecal specimens

Diagn Microbiol Infect Dis. 2014 Jan;78(1):40-4. doi: 10.1016/j.diagmicrobio.2013.09.013. Epub 2013 Sep 28.

Abstract

A new real-time reverse transcription polymerase chain reaction (RT-PCR) assay, the AccuPower Norovirus Real-time RT-PCR Kit, was evaluated in detection of human norovirus in stool specimens. Studies for detection limit, dynamic range, reproducibility, and cross-reactivity were performed. A total of 281 fecal specimens were tested using the AccuPower Norovirus Real-time RT-PCR Kit, and the results were compared with those obtained using another real-time RT-PCR system. Norovirus positivity and genotype were confirmed by direct sequencing. The lowest mean numbers of genome copies of GI and GII that could be detected by the assay were 12.3 and 5.6 RNA copies/reaction, respectively. The positive, negative, and overall percent agreements between the 2 real-time PCR assays were 99.0% (96/97), 95.1% (175/184), and 96.4% (271/281), respectively. The AccuPower Norovirus Real-time RT-PCR system showed good analytical and clinical performance and may be a useful diagnostic tool for norovirus infection.

Keywords: Comparison; Evaluation; Genotype; Norovirus; Real-time PCR.

Publication types

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

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Caliciviridae Infections / diagnosis*
  • Child
  • Child, Preschool
  • Diarrhea / diagnosis*
  • Diarrhea / virology
  • Feces / virology*
  • Female
  • Humans
  • Infant
  • Infant, Newborn
  • Male
  • Middle Aged
  • Molecular Diagnostic Techniques / methods*
  • Norovirus / isolation & purification*
  • Real-Time Polymerase Chain Reaction / methods*
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Young Adult