Development of a reverse transcription recombinase-aided amplification assay for the detection of coxsackievirus A10 and coxsackievirus A6 RNA

Arch Virol. 2018 Jun;163(6):1455-1461. doi: 10.1007/s00705-018-3734-9. Epub 2018 Feb 10.

Abstract

Hand, foot and mouth disease (HFMD) is a serious public health problem, and coxsackievirus A6 (CVA6) and coxsackievirus A10 (CVA10) are two of the major causative pathogens, in addition to enterovirus 71 (EV71) and coxsackievirus A16 (CVA16). A simple and rapid reverse transcription recombinase-aided amplification assay (RT-RAA) was developed for the detection of CVA10 and CVA6 in this study. The analytical sensitivity for detection of CVA10 and CVA6 at 95% probability by probit regression analysis was 35 copies per reaction and 38 copies per reaction, respectively, with 100% specificity. Compared with commercial RT-qPCR assays, when testing 455 fecal specimens, the kappa value of the RT-RAA assay for CVA10 and CVA6 was 0.920 (p < 0.001) and 0.952 (p < 0.001), respectively. Moreover, four samples that were positive for CVA10 and five that were positive for CVA6 by RT-RAA but negative by RT-qPCR were further determined to be true positives. These results demonstrate that the proposed RT-RAA assays are very valuable tools for the detection of CVA10 and CVA6 and have potential for use in resource-limited settings.

MeSH terms

  • Child
  • Child, Preschool
  • DNA Primers / chemistry
  • DNA Primers / genetics
  • Enterovirus / classification
  • Enterovirus / genetics*
  • Enterovirus / isolation & purification
  • Feces / virology
  • Female
  • Hand, Foot and Mouth Disease / diagnosis*
  • Hand, Foot and Mouth Disease / virology
  • Humans
  • Infant
  • Male
  • Plasmids / chemistry
  • Plasmids / metabolism
  • RNA, Viral / genetics*
  • Recombinases / chemistry*
  • Recombinases / genetics
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Reverse Transcriptase Polymerase Chain Reaction / standards
  • Sensitivity and Specificity

Substances

  • DNA Primers
  • RNA, Viral
  • Recombinases