PLoS Neglected Tropical Diseases (Sep 2016)

A Field-Deployable Reverse Transcription Recombinase Polymerase Amplification Assay for Rapid Detection of the Chikungunya Virus.

  • Pranav Patel,
  • Ahmed Abd El Wahed,
  • Oumar Faye,
  • Pauline Prüger,
  • Marco Kaiser,
  • Sasikanya Thaloengsok,
  • Sukathida Ubol,
  • Anavaj Sakuntabhai,
  • Isabelle Leparc-Goffart,
  • Frank T Hufert,
  • Amadou A Sall,
  • Manfred Weidmann,
  • Matthias Niedrig

DOI
https://doi.org/10.1371/journal.pntd.0004953
Journal volume & issue
Vol. 10, no. 9
p. e0004953

Abstract

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BACKGROUND:Chikungunya virus (CHIKV) is a mosquito-borne virus currently transmitted in about 60 countries. CHIKV causes acute flu-like symptoms and in many cases prolonged musculoskeletal and joint pain. Detection of the infection is mostly done using RT-RCR or ELISA, which are not suitable for point-of-care diagnosis. METHODOLOGY/PRINCIPAL FINDINGS:In this study, a reverse transcription recombinase polymerase amplification (RT-RPA) assay for the detection of the CHIKV was developed. The assay sensitivity, specificity, and cross-reactivity were tested. CHIKV RT-RPA assay detected down to 80 genome copies/reaction in a maximum of 15 minutes. It successfully identified 18 isolates representing the three CHIKV genotypes. No cross-reactivity was detected to other alphaviruses and arboviruses except O'nyong'nyong virus, which could be differentiated by a modified RPA primer pair. Seventy-eight samples were screened both by RT-RPA and real-time RT-PCR. The diagnostic sensitivity and specificity of the CHIKV RT-RPA assay were determined at 100%. CONCLUSIONS/SIGNIFICANCE:The developed RT-RPA assay represents a promising method for the molecular detection of CHIKV at point of need.