ORIGINAL RESEARCH article
Front. Microbiol.
Sec. Virology
Volume 16 - 2025 | doi: 10.3389/fmicb.2025.1615132
This article is part of the Research TopicViral Diagnostics: Advancements for Rapid and Enhanced DetectionView all articles
Development of a capillary-modified naked-eye visual loopmediated isothermal amplification method for the rapid detection of mpox virus and chikungunya virus
Provisionally accepted
- School of Clinical and Basic Medical Sciences, Shandong First Medical University and Shandong Academy of Medical Sciences, Jinan, China
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The global emergence of mpox virus (MPXV) and chikungunya virus (CHIKV) has intensified the demand for advanced diagnostic methods. Rapid, sensitive, costeffective diagnostic methods are crucial for preventing cross-border transmission and early containment of community spread. In this study, we developed a capillary modified Loop-mediated isothermal amplification (LAMP) assay for the identification of MPXV and CHIKV. This system employs capillaries as reaction vessels, offering advantages such as reduced reagent consumption and simplified operation. The capillary-driven liquid handling system also significantly reduces the frequency of lid openings during reagent transfer compared to standard LAMP protocols. This minimizes the risks of aerosol contamination and the associated false-positive outcomes that are inherent to conventional methods. Additionally, direct visual interpretation of the results without specialized insturmentation is achieved through integration of a Leuco-hydroxynaphthol blue (LHNB) dye. This novel detection method targets the F13 geneplasmid of MPXV, the nsP1 geneplasmid of CHIKV, live vaccinia virus (VACV) and CHIKV viruses. Analytical sensitivity reached 10 copies/μL for MPXV F13 and 6 copies/μL for CHIKV nsP1. Because of the high level of laboratory biosafety required for MPXV culture, VACV was selected as a safe surrogate model for detection, where the E9L gene was selected to target all Orthopoxvirus (OPXV). The detection limits of infectious units for intracellular and extracellular viruses of VACV are 0.64 plaque-forming units (PFU) and 8 PFU, respectively. For CHIKV infection, the detection limits of infectious units for intracellular and extracellular viruses are 0.3 PFU and 0.068 PFU, respectively. The capillary modified LAMP assay achieves higher sensitivity to current gold-standard qPCR assays, while offering several advantages, including rapid turnaround time (results obtained within 30 minutes), minimal equipment requirements (single heating module), cost-effectiveness, visual readout compatibility, and no requirement for specialized personnel. This study confirmed the capacity of this improved LAMP colorimetric detection method. The system addresses critical gaps in resource-limited scenarios, offering a deployable solution for border quarantine stations and primary healthcare services--key nodes for intercepting cross-border transmission and mitigating localized outbreaks through timely case identification.
Keywords: LAMP, capillary, MPXV, CHIKV, diagnostic method
Received: 20 Apr 2025; Accepted: 16 Jul 2025.
Copyright: © 2025 Luan, 宋, Cheng, Li, Zhu, Cheng and Zhang. 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: Leiliang Zhang, School of Clinical and Basic Medical Sciences, Shandong First Medical University and Shandong Academy of Medical Sciences, Jinan, China
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