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

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 poi...

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Bibliographic Details
Published in:PLOS Neglected Tropical Diseases
Main Authors: 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
Format: Article in Journal/Newspaper
Language:English
Published: Public Library of Science (PLoS) 2016
Subjects:
Arctic medicine. Tropical medicine
RC955-962
Public aspects of medicine
RA1-1270
Arctic
Online Access:https://doi.org/10.1371/journal.pntd.0004953
https://doaj.org/article/9477b88df5c0426ab96f914053308d89
Description
Summary: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.

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