A field-deployable reverse transcription recombinase polymerase amplification assay for rapid detection of the Chikungunya virus

Citation

Patel P, El Wahed AA, Faye O, Pruger P, Kaiser M, Thaloengsok S, Ubol S, Sakuntabhai A, Leparc-Goffart I, Hufert FT, Sall AA, Weidmann M & Niedrig M (2016) A field-deployable reverse transcription recombinase polymerase amplification assay for rapid detection of the Chikungunya virus. PLoS Neglected Tropical Diseases, 10 (9), Art. No.: e0004953. https://doi.org/10.1371/journal.pntd.0004953

Abstract
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.

Journal
PLoS Neglected Tropical Diseases: Volume 10, Issue 9