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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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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5042537PMC
http://dx.doi.org/10.1371/journal.pntd.0004953DOI Listing

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  • - The study focuses on creating a rapid and sensitive diagnostic test for Chikungunya virus (CHIKV) using a combination of CRISPR/Cas12a technology and reverse transcription recombinase polymerase amplification (RT-RPA) to address underdiagnosis and reduce hospital burdens caused by delayed or incorrect diagnoses.
  • - The new assay can detect CHIKV within 35 minutes and allows dual detection methods—both fluorescence and lateral flow—without opening the reaction tube, minimizing contamination risks.
  • - It boasts a high level of sensitivity and specificity, accurately detecting as low as 8 gene copies of CHIKV, while avoiding cross-reactivity with other related pathogens, making it a promising tool for quick clinical diagnosis
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