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Development and proof-of-concept evaluation for a low resource compatible Chikungunya virus diagnostic.

Rickyle Balea1,2,3, Alberto A Amarilla2, Jody Hobson-Peters2

  • 1School of Science, Technology and Engineering, University of the Sunshine Coast, Sippy Downs, Queensland, Australia.

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Summary

A new diagnostic test using reverse transcription-recombinase aided amplification (RT-RAA) offers rapid and sensitive detection of Chikungunya virus (CHIKV). This point-of-care test is faster and more accessible than traditional methods, aiding in early CHIKV outbreak mitigation.

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Area of Science:

  • Virology
  • Molecular Diagnostics
  • Public Health

Background:

  • Chikungunya virus (CHIKV) poses a significant global health threat, transmitted primarily by Aedes mosquitoes.
  • Current diagnostic and therapeutic options for CHIKV are limited, emphasizing the need for early detection.
  • Existing diagnostic methods like RT-qPCR are time-consuming and require specialized equipment.

Purpose of the Study:

  • To develop and evaluate a sensitive and specific diagnostic tool for Chikungunya virus detection.
  • To create a rapid, point-of-care diagnostic assay for CHIKV RNA.
  • To offer an alternative to conventional CHIKV diagnostic methods.

Main Methods:

  • Designed a diagnostic assay combining reverse transcription-recombinase aided amplification (RT-RAA) with lateral flow strip detection (LFD).
  • Targeted a conserved region of the CHIKV E1 gene for detection.
  • Utilized a novel sample preparation reagent (TNA-Cifer-E) for CHIKV inactivation and RNA preservation.

Main Results:

  • The developed assay (Iso-CHIKV-Dx) demonstrated high sensitivity and specificity, detecting CHIKV RNA in urine samples.
  • The assay successfully inactivated live CHIKV within two minutes at room temperature while preserving RNA integrity.
  • Iso-CHIKV-Dx detected as low as 570 copies/µL of CHIKV RNA in 30 minutes under isothermal conditions.
  • The diagnostic showed no cross-reactivity with related Alphaviruses or common Flaviviruses.
  • The assay proved to be four times faster than conventional RNA isolation and RT-qPCR methods.

Conclusions:

  • The Iso-CHIKV-Dx is a robust, rapid, and sensitive point-of-care diagnostic for Chikungunya virus.
  • This assay offers a valuable alternative to conventional diagnostics, especially in resource-limited settings.
  • The developed diagnostic has the potential to significantly aid in the early detection and management of CHIKV outbreaks.