Korean Journal of Veterinary Service (한국동물위생학회지)

The Korean Society of Veterinary Service (한국동물위생학회)
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Simple and rapid colorimetric detection of African swine fever virus by loop-mediated isothermal amplification assay using a hydroxynaphthol blue metal indicator

  • Park, Ji-Hoon (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University) ;
  • Kim, Hye-Ryung (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University) ;
  • Chae, Ha-Kyung (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University) ;
  • Park, Jonghyun (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University) ;
  • Jeon, Bo-Young (Department of Biomedical Laboratory Science, College of Health Science, Yonsei University) ;
  • Lyoo, Young S. (College of Veterinary Medicine, Konkuk University) ;
  • Park, Choi-Kyu (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University)
  • Received : 2022.03.23
  • Accepted : 2022.03.28
  • Published : 2022.03.30
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Abstract

In this study, a simple loop-mediated isothermal amplification (LAMP) combined with visual detection method (vLAMP) assay was developed for the rapid and specific detection of African swine fever virus (ASFV), overcoming the shortcomings of previously described LAMP assays that require additional detection steps or pose a cross-contamination risk. The assay results can be directly detected by the naked eye using hydroxynaphthol blue after incubation for 40 min at 62℃. The assay specifically amplified ASFV DNA and no other viral nucleic acids. The limit of detection of the assay was <50 DNA copies/reaction, which was ten times more sensitive than conventional polymerase chain reaction (cPCR) and comparable to real-time PCR (qPCR). For clinical evaluation, the ASFV detection rate of vLAMP was higher than cPCR and comparable to OIE-recommended qPCR, showing 100% concordance, with a κ value (95% confidence interval) of 1 (1.00~1.00). Considering the advantages of high sensitivity and specificity, no possibility for cross-contamination, and being able to be used as low-cost equipment, the developed vLAMP assay will be a valuable tool for detecting ASFV from clinical samples, even in resource-limited laboratories.

Keywords

Acknowledgement

This work was supported by the Commercializations Promotion Agency for R&D Outcomes (COMPA) grant funded by the Korean Government(Ministry of Science and ICT) (2022).

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