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Journal of General Virology header logo Journal of General Virology

Volume 106, Issue 5

Development of a loop-mediated isothermal amplification assay for the rapid detection of Alongshan virus Open Access

Abstract

Alongshan virus (ALSV) is a recently discovered tick-borne zoonotic virus. Currently, there is no rapid and accurate clinical method for ALSV detection. This study aimed to develop a loop-mediated isothermal amplification (LAMP) assay for precise ALSV infection detection. Specific primers were designed based on the S1 segment of the ALSV NE-TH4 strain’s genome (GenBank accession no. ON408067.1). The reaction time, temperature and concentration of the neutral red staining solution in the LAMP assay were optimized. Thorough evaluations of specificity, sensitivity and repeatability led to the development of a visually interpretable LAMP assay. The optimal amplification time was 50 min. The minimum detection limit for cDNA was as low as 0.005 pg μl, and sensitivity for standards was 1.68×10 copies per μl, surpassing that of PCR and real-time PCR. No cross-reactivity was observed with Jingmen tick virus, Bole tick virus 4 and Beiji nairovirus. These results indicate that the LAMP assay is more sensitive and accurate than PCR and real-time PCR. The developed LAMP assay allows for on-site detection, reduces testing costs and provides rapid and accurate results. Thus, it lays a solid foundation for the prevention and control of emerging tick-borne ALSV.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): Alongshan virus (ALSV) , loop-mediated isothermal amplification technique (LAMP) , PCR , rapid detection and visualization
Funding
This study was supported by the:
  • the Science and Technology Project in Guangzhou Province (Award 202103000008-1)
    • Principle Award Recipient: LiuQuan
  • Cultivating Plan Program for the Leader in Science and Technology of Yunnan Province (Award 2019CX01N111)
    • Principle Award Recipient: LiuQuan
  • the Research Capacity Improvement Project of Key Disciplines in Guangdong Province (Award 2021ZDJS100)
    • Principle Award Recipient: LiuQuan
© 2025 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. The Microbiology Society waived the open access fees for this article.
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2025-05-08
2025-05-18
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/content/journal/jgv/10.1099/jgv.0.002094
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Development of a loop-mediated isothermal amplification assay for the rapid detection of Alongshan virus
J. Gen. Virol. 106, 002094 (2025); https://doi.org/10.1099/jgv.0.002094
/content/journal/jgv/10.1099/jgv.0.002094
/content/journal/jgv/10.1099/jgv.0.002094
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