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Purpose <p>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.</p> Methods <p>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.</p> Results <p>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; <i>p</i> &lt; 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 (<i>κ</i> = 1.00; 95% CI: 1.00–1.00). The actual detection time was ≤ 20&#xa0;min, and the assay performed reliably in low-viral-load and co-infection cases.</p> Conclusion <p>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.</p>

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

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年8月5日

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  • Mengjie Liang,
  • Chunyan Liu,
  • Yongxin Li,
  • Qian Shi,
  • Hefei Zha,
  • Jingyuan Yang,
  • Zhaohui Deng,
  • Xin Zhang

摘要

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.