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

Pranav Patel (1) , Ahmed Abd El Wahed (2) , Oumar Faye (3) , Pauline Prüger (1) , Marco Kaiser (4) , Sasikanya Thaloengsok (5) , Sukathida Ubol (5) , Anavaj Sakuntabhai (6) , Isabelle Leparc-Goffart (7) , Frank T Hufert (8) , Amadou A Sall (3) , Manfred Weidmann (9) , Matthias Niedrig (1)

1 RKI - Robert Koch Institute [Berlin]
2 Georg-August-University = Georg-August-Universität Göttingen
3 Arbovirus et Virus de Fièvres Hémorragiques [Dakar, Sénégal]
4 GenExpress Gesellschaft für Proteindesign mbH
5 Mahidol University [Bangkok]
6 Génétique fonctionnelle des Maladies infectieuses - Functional Genetics of Infectious Diseases
7 IRBA - Institut de Recherche Biomédicale des Armées [Antenne Marseille]
8 MHB Theodor Fontane - Medizinische Hochschule Brandenburg Theodor Fontane / Brandenburg Medical School Theodor-Fontane
9 University of Stirling

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.

Domains: Virology; Biochemistry, Molecular Biology

Pranav Patel, Ahmed Abd El Wahed, Oumar Faye, Pauline Prüger, Marco Kaiser, et al.. A Field-Deployable Reverse Transcription Recombinase Polymerase Amplification Assay for Rapid Detection of the Chikungunya Virus. PLoS Neglected Tropical Diseases, 2016, 10 (9), pp.e0004953. ⟨10.1371/journal.pntd.0004953⟩. ⟨pasteur-02083456⟩