Liang Mengjie, Liu Chunyan, Li Yongxin, Shi Qian, Zha Hefei, Yang Jingyuan, Deng Zhaohui, Zhang Xin
Department of Medical Laboratory, Hospital of Xinjiang Production and Construction Corps/Second Affiliated Hospital of Shihezi University School of Medicine, No. 232, Qingnian Road, Tianshan District, Urumqi, 830002, Xinjiang, China.
Bingtuan Clinical Medical Research Center for Medical Laboratory, No. 232, Qingnian Road, Tianshan District, Urumqi, 830002, Xinjiang, China.
Eur J Clin Microbiol Infect Dis. 2025 Aug 5. doi: 10.1007/s10096-025-05224-3.
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.
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.
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.
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.
我们开发了一种环介导等温扩增（LAMP）多平台检测系统，该系统克服了传统聚合酶链反应（PCR）在及时性和设备依赖性方面的局限性，为不同资源条件的临床环境建立了一种快速、准确诊断人腺病毒3型和7型（HAdV-3和HAdV-7）的框架。
基于HAdV-3和HAdV-7六邻体基因的保守区域设计LAMP引物。采用钙黄绿素法、免疫层析法（IC）和荧光探针法。通过敏感性和特异性分析确定每种方法的性能。此外，使用商业PCR试剂盒对188份临床样本进行平行分析。
钙黄绿素法和IC法的检测限（LOD）为2.5拷贝/反应。荧光探针法表现出更高的敏感性，LOD为1拷贝/反应，中位Ct值为7.3，比定量PCR（qPCR）的中位Ct值26.9低72.8%（p<0.05）。所有三个平台的特异性均为100%，未观察到与严重急性呼吸综合征冠状病毒2（SARS-CoV-2）或其他检测的呼吸道病原体发生交叉反应。临床验证显示荧光探针LAMP检测法与qPCR的一致性为100%（κ=1.00；95%CI：1.00-1.00）。实际检测时间≤20分钟，该检测法在低病毒载量和合并感染病例中表现可靠。
本研究建立的多平台LAMP系统构建了一个“初步筛查-定量”的分级检测网络，专门设计用于满足基层、现场和实验室环境的多样化需求。该系统为呼吸道感染的防控提供了高效的多场景解决方案。
