METHODS article
Front. Microbiol.
Sec. Infectious Agents and Disease
Volume 16 - 2025 | doi: 10.3389/fmicb.2025.1678396
This article is part of the Research TopicRapid and Efficient Analytical Technologies for Pathogen DetectionView all 21 articles
A Rapid and Visual Dual LAMP-LFD Assay for On-Site Simultaneous Detection of Influenza A Virus (H1N1) and Respiratory Syncytial Virus (RSV)
Provisionally accepted
Alang Zhang1Xingyu Lan2Hengxuan Zhou1Ting Zhou1Zhihua Xu1Xinyu Cheng1Chenxi Guo1Mingjie Wei1Jiahui Wu1
Shi Feng1*- 1College of Life Sciences, Shihezi University, Shihezi, China
- 2The First Division Hospital of Xinjiang Production and Construction Corps, Akesu, China
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Respiratory tract infections caused by influenza A virus (IAV) and respiratory syncytial virus (RSV) are a major global health burden. To overcome the limitations of existing diagnostic techniques for timely detection (point-of-care testing), this study developed a dual rapid visual detection method based on loop-mediated isothermal amplification (LAMP) and lateral flow device (LFD) technologies for the simultaneous detection of H1N1 influenza virus and RSV. This method uses a dual-labeled probe system (H1N1: digoxigenin/biotin; RSV: 6-carboxyfluorescein/biotin) combined with a two-color latex microsphere signal system that enables the intuitive visual interpretation of multiple detection results. Compared with traditional nucleic acid detection, the entire detection process was completed within 40 min at a constant temperature of 63 °C and the operation was simple. After optimization, the method showed good sensitivity and specificity. The limit of detection for H1N1 IAV and RSV was as low as 7.78 × 103 copies/mL and 1.29 × 102 copies/mL, respectively, and no cross-reaction occurred with six common non-target pathogens. Clinical sample verification showed that the consistency between the detection and RT-qPCR results was 96.83 %, which was significantly better than traditional antigen detection. The dual LAMP-LFD detection method established in this study provides an efficient solution for rapid on-site, synchronous, and visual screening in resource-limited environments and has important clinical application value.
Keywords: double detection, respiratory viruses, Loop-mediated isothermal amplification, Lateral flow device, visual detection
Received: 02 Aug 2025; Accepted: 21 Oct 2025.
Copyright: © 2025 Zhang, Lan, Zhou, Zhou, Xu, Cheng, Guo, Wei, Wu and Feng. 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: Shi Feng, shifeng2314@yeah.net
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