Development of Nucleic Acid Lateral Flow Immunoassay for Rapid and Accurate Detection of Chikungunya Virus in Indonesia

J Microbiol Biotechnol. 2021 Dec 28;31(12):1716-1721. doi: 10.4014/jmb.2108.08025.

Abstract

Chikungunya fever is an arboviral disease caused by the Chikungunya virus (CHIKV). The disease has similar clinical manifestations with other acute febrile illnesses which complicates differential diagnosis in low-resource settings. We aimed to develop a rapid test for CHIKV detection based on the nucleic acid lateral flow immunoassay technology. The system consists of a primer set that recognizes the E1 region of the CHIKV genome and test strips in an enclosed cassette which are used to detect amplicons labeled with FITC/biotin. Amplification of the viral genome was done using open-source PCR, a low-cost open-source thermal cycler. Assay performance was evaluated using a panel of RNA isolated from patients' blood with confirmed CHIKV (n = 8) and dengue virus (n = 20) infection. The open-source PCR-NALFIA platform had a limit of detection of 10 RNA copies/ml. The assay had a sensitivity and specificity of 100% (95% CI: 67.56% - 100%) and 100% (95% CI: 83.89% - 100%), respectively, compared to reference standards of any positive virus culture on C6/36 cell lines and/or qRT-PCR. Further evaluation of its performance using a larger sample size may provide important data to extend its usefulness, especially its utilization in the peripheral healthcare facilities with scarce resources and outbreak situations.

Keywords: Chikungunya fever; Indonesia; NALFIA; clinical validation; open-source PCR.

MeSH terms

  • Chikungunya Fever / blood
  • Chikungunya Fever / diagnosis*
  • Chikungunya virus / genetics
  • Chikungunya virus / isolation & purification*
  • Genome, Viral / genetics
  • Humans
  • Immunoassay
  • Indonesia
  • Limit of Detection
  • Molecular Diagnostic Techniques / instrumentation
  • Molecular Diagnostic Techniques / methods*
  • Molecular Diagnostic Techniques / standards
  • Polymerase Chain Reaction
  • RNA, Viral / blood
  • RNA, Viral / genetics
  • Sensitivity and Specificity

Substances

  • RNA, Viral