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ORIGINAL RESEARCH article

Front. Microbiol.
Sec. Microbiotechnology
Volume 15 - 2024 | doi: 10.3389/fmicb.2024.1403577

Ultra-rapid and sensitive detection of African swine fever virus using multiple cross displacement amplification combined with nanoparticle-based lateral flow biosensor

Provisionally accepted
Sha Mao Sha Mao 1Renjun Zhang Renjun Zhang 2Xinggui Yang Xinggui Yang 3Junfei Huang Junfei Huang 3Yingqian Kang Yingqian Kang 4Yi Wang Yi Wang 5Hong Chen Hong Chen 6Shijun Li Shijun Li 3*
  • 1 Guizhou Centre for Disease Control and Prevention, Guiyang, China
  • 2 Guizhou Provincial Center for Animal Disease Control and Prevention, Guiyang, China
  • 3 Guizhou Provincial Center for Disease Control and Prevention, Guiyang, China
  • 4 Guizhou Medical University, Guiyang, Guizhou Province, China
  • 5 Experimental Research Center, Capital Institute of Pediatrics, Beijing, China
  • 6 EPINTEK Guiyang Ltd, Guiyang, Guizhou Province, China

The final, formatted version of the article will be published soon.

    African swine fever (ASF) is a devastating disease that can kill almost all infected pigs, causing great damage to the pig industry and destabilizing the global economy.Here, we developed a specific assay that combined multiple cross-displacement amplification (MCDA) with a nanoparticle-based lateral flow biosensor (LFB) for early and rapid identification of the African swine fever virus (ASFV-MCDA-LFB).We first designed a set of MCDA primers to recognize ten different regions of the target ASFV B646L gene. Subsequently, the MCDA reaction was monitored with various methods: MG chromogenic reagents, agarose gel electrophoresis, real-time turbidity, and LFB. The ASFV-MCDA-LFB assay was optimized and evaluated with target nucleic acid templates extracted from various pathogens and simulated whole blood samples. As a result, the detection of limit (LOD) of the ASFV assay was 200 copies/reaction within 30 minutes, and no cross-reaction were observed with other non-ASFV viruses and common pathogens in this study. The evaluation assays demonstrated that the ASFV-MCDA-LFB method here is rapid, objective, easy-to-use, and low-cost detection method which can be used as a diagnostic or screening tool with competitive potential for point-of-care testing (POCT) of ASFV.

    Keywords: African swine fever virus1, multiple cross displacement amplification2, Nanoparticle-based lateral flow biosensor3, ASFV-MCDA-LFB4, Point-of-care Testing5

    Received: 19 Mar 2024; Accepted: 28 Oct 2024.

    Copyright: © 2024 Mao, Zhang, Yang, Huang, Kang, Wang, Chen and Li. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Shijun Li, Guizhou Provincial Center for Disease Control and Prevention, Guiyang, China

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.