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
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.
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%.
The developed RT-RPA assay represents a promising method for the molecular detection of CHIKV at point of need.
PLoS Neglected Tropical Diseases: Volume 10, Issue 9
|Publication date online||29/09/2016|
|Date accepted by journal||03/08/2016|
|Publisher||Public Library of Science|