Publications
Microchemical JournalApr 2025 |
213
113763-113763
DOI:
10.1016/j.microc.2025.113763

Development of triplex loop-mediated isothermal amplification with immunochromatographic assay for rapid simultaneous detection three respiratory viruses

Liu, Jie; Sun, Rui; Ru, Jing; Ma, Wenxin; Shen, Qiang; Chen, Mingli; Yin, Jian; Ma, Fuqiang; Yin, Huancai
Product Used
Genes
Abstract
Influenza is the leading cause of human respiratory infections and poses a significant threat to human health. Given the genetic diversity of influenza viruses, it is crucial to develop high-throughput assays capable of simultaneously detecting multiple influenza strains in a single reaction. We developed a rapid, sensitive, and visual point-of-care detection method for three respiratory viruses (H1N1, H3N2, IBV) in throat swabs by integrating multiplex loop-mediated isothermal amplification (LAMP) with lateral flow immunoassay (LFIA). In this study, the LAMP primers for influenza A (H1N1 and H3N2) and influenza B viruses (IBV) were labeled with Texas Red, biotin, and FAM, respectively. The LAMP amplification products containing different markers were captured by corresponding antibodies on the LFIA strip, forming visible bands. The results demonstrated that the triplex LAMP assay showed high sensitivity, with a limit of detection (LOD) of 0.2 copies/μL for both H1N1 and IBV, and 20 copies/μL for H3N2, as well as high specificity achieved by optimizing the primer ratio. The integration of uracil DNA glycosylase effectively mitigates carryover contamination in LAMP, addressing the inherent contradiction caused by uncapping in the lateral flow strategy, while maintaining isothermal conditions throughout the process. Furthermore, the entire detection process can be completed within 40 min. Therefore, the triplex LAMP-LFIA shows great potential and a promising future for the detection of influenza viruses in clinical samples.
Product Used
Genes

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