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

作者：Patel Pranav; Abd El Wahed Ahmed; Faye Oumar; Prueger Pauline; Kaiser Marco; Thaloengsok Sasikanya; Ubol Sukathida; Sakuntabhai Anavaj; Leparc Goffart Isabelle; Hufert Frank T; Sall Amadou A; Weidmann Manfred; Niedrig Matthias^*

来源：PLoS Neglected Tropical Diseases, 2016, 10(9): e0004953.

DOI：10.1371/journal.pntd.0004953

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.

• 出版日期2016-9