Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Development of Nucleic Acid Lateral Flow Immunoassay for Rapid and Accurate Detection of Chikungunya Virus in Indonesia

  • Ajie, Mandala (Research Center for Care and Control of Infectious Disease (RC3ID), Faculty of Medicine, Universitas Padjadjaran) ;
  • Pascapurnama, Dyshelly Nurkartika (Research Center for Care and Control of Infectious Disease (RC3ID), Faculty of Medicine, Universitas Padjadjaran) ;
  • Prodjosoewojo, Susantina (Research Center for Care and Control of Infectious Disease (RC3ID), Faculty of Medicine, Universitas Padjadjaran) ;
  • Kusumawardani, Shinta (Research Center for Care and Control of Infectious Disease (RC3ID), Faculty of Medicine, Universitas Padjadjaran) ;
  • Djauhari, Hofiya (Research Center for Care and Control of Infectious Disease (RC3ID), Faculty of Medicine, Universitas Padjadjaran) ;
  • Handali, Sukwan (Division of Parasitic Diseases and Malaria, Centers for Disease Control and Prevention) ;
  • Alisjahbana, Bachti (Research Center for Care and Control of Infectious Disease (RC3ID), Faculty of Medicine, Universitas Padjadjaran) ;
  • Chaidir, Lidya (Research Center for Care and Control of Infectious Disease (RC3ID), Faculty of Medicine, Universitas Padjadjaran)
  • Received : 2021.08.19
  • Accepted : 2021.09.29
  • Published : 2021.12.28
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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

Acknowledgement

We would like to thank our colleagues in the Infectious Disease Research Center for their support in getting the study done and for a fruitful discussion during the study period. This study was supported by the Grand Challenge Canada and Universitas Padjadjaran. The funding sources had no role in the design, conduct, analysis or reporting of the study.

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