Abstract
BackgroundChikungunya 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 FindingsIn 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/SignificanceThe developed RT-RPA assay represents a promising method for the molecular detection of CHIKV at point of need.
Highlights
Chikungunya virus (CHIKV) is a mosquito-borne virus belonging to the genus alphavirus, family Togaviridae and spread by Aedes mosquitoes
CHIKV is transmitted to humans via mosquitos
We have developed a molecular assay for the detection of CHIKV genome based on isothermal„recombinase polymerase amplification (RPA) assay”performed at 42°C
Summary
Chikungunya virus (CHIKV) is a mosquito-borne virus belonging to the genus alphavirus, family Togaviridae and spread by Aedes mosquitoes. CHIKV is the aetiological agent of chikungunya fever (CF) and was first isolated in 1952 from the serum of a febrile patient during an outbreak in Tanzania, Africa [1]. CHIKV infection is characterized by an abrupt onset of fever lasting two to five days, frequently accompanied by arthralgia. Other common symptoms include muscle pain, headache, nausea, fatigue and rash similar to the dengue virus infection. Besides CHIKV, the Alphavirus genus includes viruses such as O'nyong'nyong virus (ONNV), Ross River virus (RRV), Barmah Forest virus (BFV), Sindbis virus (SNV) and Mayaro virus (MAYV) [5]. 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
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