A new accurate assay for Coxsackievirus A 16 by fluorescence detection of isothermal RNA amplification

J Virol Methods. 2013 Nov;193(2):459-62. doi: 10.1016/j.jviromet.2013.07.013. Epub 2013 Jul 18.

Abstract

Coxsackievirus A 16 (CA16) is one of the most common causes of hand, foot, and mouth disease (HFMD) worldwide. Without a vaccine or antiviral drug early, rapid, and accurate detection is critical for preventing and controlling HFMD. A simultaneous amplification and testing (SAT) assay was developed for detecting CA16 based on isothermal RNA amplification with fluorescence using standard, real-time PCR equipment. Primers and probes were designed to target the VP1 region of CA16. Virus strains and clinical specimens were used to evaluate the diagnostic performance characteristics of the assay. The assay detected as few as 10 copies of CA16 RNA transcripts. Using real-time PCR plus sequencing as the reference standard, the sensitivity and specificity of the SAT-CA16 assay were 100% and 99.2%, respectively. These findings indicate that SAT-CA16 is a rapid and reliable method for detecting CA16.

Keywords: CA16; Isothermal RNA amplification; Simultaneous amplification and testing (SAT).

Publication types

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

MeSH terms

  • Child, Preschool
  • DNA Primers / genetics
  • Enterovirus / genetics
  • Enterovirus / isolation & purification*
  • Fluorescence
  • Hand, Foot and Mouth Disease / diagnosis*
  • Hand, Foot and Mouth Disease / virology
  • Humans
  • Molecular Diagnostic Techniques / methods*
  • RNA, Viral / genetics
  • Real-Time Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Viral Structural Proteins / genetics
  • Virology / methods

Substances

  • DNA Primers
  • RNA, Viral
  • Viral Structural Proteins