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

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...

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Bibliographic Details
Published in:	PLoS neglected tropical diseases Vol. 10; no. 9; p. e0004953
Main Authors:	Patel, Pranav, Abd El Wahed, Ahmed, Faye, Oumar, Prüger, Pauline, Kaiser, Marco, Thaloengsok, Sasikanya, Ubol, Sukathida, Sakuntabhai, Anavaj, Leparc-Goffart, Isabelle, Hufert, Frank T, Sall, Amadou A, Weidmann, Manfred, Niedrig, Matthias
Format:	Journal Article
Language:	English
Published:	United States Public Library of Science 29-09-2016 Public Library of Science (PLoS)
Subjects:	Biochemistry, Molecular Biology Biology and life sciences Chikungunya virus Dengue fever Funding Gene amplification Genetic aspects Genomes Genotype & phenotype Genotypes Health aspects Infections Life Sciences Medicine and Health Sciences Methods Microbiological assay Microbiology and Parasitology Pain Research and Analysis Methods Tropical diseases Vector-borne diseases Virology Zika virus Thailand Bangkok Thailand
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Summary:	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. 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%. The developed RT-RPA assay represents a promising method for the molecular detection of CHIKV at point of need.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 PMCID: PMC5042537 All authors have no financial interests except MK is employed by GenExpress Gesellschaft für Proteindesign and has commercial interest in the molecular RNA standard. This does not alter the authors´ adherence to all PLOS policies on sharing data and materials. Conceived and designed the experiments: PPa AAEW OF MW MN.Performed the experiments: PPa AAEW OF PPr MK ST SU AS ILG FTH AAS MW MN.Analyzed the data: PPa AAEW OF PPr MK ST SU AS ILG FTH AAS MW MN.Contributed reagents/materials/analysis tools: OF SU ILG AAS.Wrote the paper: PPa AAEW MW MN.Critical revision of the manuscript: PPa AAEW OF PPr MK ST SU AS ILG FTH AAS MW MN.
ISSN:	1935-2735 1935-2727 1935-2735
DOI:	10.1371/journal.pntd.0004953