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. 2025 Aug 5.
doi: 10.1007/s10096-025-05224-3. Online ahead of print.

Development and validation of a multi-platform LAMP system for rapid HAdV-3 and HAdV-7 detection

Mengjie Liang  1   2 , Chunyan Liu  1   2 , Yongxin Li  1   2 , Qian Shi  1   2 , Hefei Zha  1   2 , Jingyuan Yang  1   2 , Zhaohui Deng  3   4 , Xin Zhang  5   6
Affiliations

Development and validation of a multi-platform LAMP system for rapid HAdV-3 and HAdV-7 detection

Mengjie Liang et al. Eur J Clin Microbiol Infect Dis. .

Abstract

Purpose: We developed a loop-mediated isothermal amplification (LAMP) multi-platform detection system that overcomes the limitations of traditional polymerase chain reaction (PCR) in timeliness and equipment dependency, establishing a rapid, accurate diagnostic framework for Human adenovirus types 3 and 7 (HAdV-3 and HAdV-7) across resource-varied clinical settings.

Methods: LAMP primers were designed based on the conserved regions of the Hexon genes of HAdV-3 and HAdV-7. The calcein, immunochromatography (IC), and fluorescence probe methods were used. Sensitivity and specificity analyses determined each method's performance. Additionally, 188 clinical samples were analyzed in parallel using a commercial PCR kit.

Results: The calcein and IC methods achieved a limit of detection (LOD) of 2.5 copies/reaction. The fluorescent probe method demonstrated superior sensitivity, with an LOD of 1 copy/reaction and a median Ct value of 7.3, 72.8% lower than that of qPCR (median Ct 26.9; p < 0.05). All three platforms exhibited 100% specificity, with no cross-reactivity observed against SARS-CoV-2 or other tested respiratory pathogens. Clinical validation showed 100% concordance between the fluorescent probe LAMP assay and qPCR (κ = 1.00; 95% CI: 1.00-1.00). The actual detection time was ≤ 20 min, and the assay performed reliably in low-viral-load and co-infection cases.

Conclusion: The multi-platform LAMP system established in this study has created a hierarchical detection network characterized as "preliminary screening-quantitative", specifically designed to meet the diverse needs of grassroots, field, and laboratory settings. This system offers efficient multi-scenario solutions for the prevention and control of respiratory infections.

Keywords: Calcein; HAdV; Immunochromatography; LAMP; Respiratory infections.

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Conflict of interest statement

Declarations. Ethics approval: This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the Ethics Committee of the Xinjiang Production and Construction Corps Hospital (Approval No 202206301). Consent to participate: The need for informed consent was waived due to the retrospective use of anonymized samples. Consent to publish: Not applicable. Competing interests: The authors declare no competing interests.

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