Cui Jinling, Shi Binghui, Chai Mengnan, Wang Yanling, Du Jiansen, Lu Xiaoguang, Shi Chao, Ma Cuiping, Li Yong
College of Biological Engineering, Sino-UAE International Cooperative Joint Laboratory of Pathogenic Microorganism Rapid Detection, Qingdao Key Laboratory of Rapid Nucleic Acid Detection, Qingdao Rapid Nucleic Acid Detection Engineering Research Center, Qingdao University of Science and Technology, Qingdao, 266042, China.
Qingdao International Travel Healthcare Center, Qingdao Customs District, Qingdao, 266071, China.
Mikrochim Acta. 2025 Aug 6;192(9):559. doi: 10.1007/s00604-025-07377-6.
Mycoplasma pneumoniae is a major cause of community-acquired pneumonia, yet its diagnosis remains challenging due to overlapping symptoms with other respiratory pathogens. In this study, an integrated microfluidic cartridge platform has been developed that combines three synergistic advancements: a chemical lysis reagent (GuHCl/EDTA) enabling direct genomic DNA release from M. pneumoniae collected by nasal swabs within 2 min without extraction steps, a novel dual-dye system (hydroxy naphthol blue/SYBR Green I) that leverages complementary fluorescence transitions from red to green for enhanced visual discrimination of loop-mediated isothermal amplification (LAMP) results, and a smartphone-based RGB quantification algorithm to standardize interpretation by analyzing colorimetric changes. The microfluidic cartridge integrates lysis, amplification, and detection into a closed workflow, achieving a sample-to-answer time of 40 min with a detection limit of 1 copy/μL. Clinical validation using 25 nasal swab samples demonstrated 95% sensitivity (19/20) and 100% specificity (5/5) compared with serological tests. This platform offers a rapid, cost-effective, and user-friendly solution for point-of-care diagnosis of M. pneumoniae, particularly in resource-limited settings.
肺炎支原体是社区获得性肺炎的主要病因，但由于其症状与其他呼吸道病原体重叠，其诊断仍然具有挑战性。在本研究中，开发了一种集成微流控芯片平台，该平台结合了三项协同进展：一种化学裂解试剂（盐酸胍/乙二胺四乙酸），可在2分钟内从鼻拭子采集的肺炎支原体中直接释放基因组DNA，无需提取步骤；一种新型双染料系统（羟基萘酚蓝/ SYBR Green I），利用从红色到绿色的互补荧光转变来增强对环介导等温扩增（LAMP）结果的视觉辨别；以及一种基于智能手机的RGB定量算法，通过分析比色变化来标准化解释。该微流控芯片将裂解、扩增和检测集成到一个封闭的工作流程中，实现了40分钟的样本到答案时间，检测限为1拷贝/微升。使用25份鼻拭子样本进行的临床验证表明，与血清学检测相比，灵敏度为95%（19/20），特异性为100%（5/5）。该平台为肺炎支原体的即时诊断提供了一种快速、经济高效且用户友好的解决方案，特别是在资源有限的环境中。
