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 point-of-care diagnosis.


RESULTS
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
The developed RT-RPA assay represents a promising method for the molecular detection of CHIKV at point of need.
Centre for Biological Threats and Special Pathogens, Robert Koch Institute, Berlin, Germany.
Pranav
P
Patel
Division of Microbiology and Animal Hygiene, Georg-August-University, Goettingen, Germany.
Ahmed
A
Abd El Wahed
Arbovirus Unit, Pasteur Institute, Dakar, Senegal.
Oumar
O
Faye
Centre for Biological Threats and Special Pathogens, Robert Koch Institute, Berlin, Germany.
Pauline
P
Prüger
GenExpress Gesellschaft für Proteindesign, Berlin, Germany.
Marco
M
Kaiser
Department of Microbiology, Faculty of Science, Mahidol University, Bangkok, Thailand.
Sasikanya
S
Thaloengsok
Department of Microbiology, Faculty of Science, Mahidol University, Bangkok, Thailand.
Sukathida
S
Ubol
Functional Genetics of Infectious Diseases Unit, Institute Pasteur, Paris, France.
Anavaj
A
Sakuntabhai
French Armed Force Biomedical Research Institute (IRBA) Marseille, France.
Isabelle
I
Leparc-Goffart
Institute of Microbiology & Virology, Brandenburg Medical School Fontane, Senftenberg, Germany.
Frank T
FT
Hufert
Arbovirus Unit, Pasteur Institute, Dakar, Senegal.
Amadou A
AA
Sall
Institute of Aquaculture, University of Stirling, Stirling, Scotland, United Kingdom.
Manfred
M
Weidmann
Centre for Biological Threats and Special Pathogens, Robert Koch Institute, Berlin, Germany.
Matthias
M
Niedrig
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