Jiang Tingqing, Wei Guijiang, Lin Mei, Zhang Sheping, Zou Lintao, Zhou Xuan, Deng Zhongliang
Affiliated Hospital of Xiangnan University, Chenzhou, 423099, China.
Center for Medical Laboratory Science, Affiliated Hospital of Youjiang Medical University for Nationalities, Guangxi, 533000, China; Baise Key Laboratory for Research and Development on Clinical Molecular Diagnosis for High-Incidence Diseases, Guangxi, 533000, China.
Talanta. 2025 May 15;287:127615. doi: 10.1016/j.talanta.2025.127615. Epub 2025 Jan 24.
Rapid and accurate detection of Chlamydia psittaci, the causative agent of psittacosis, is crucial for both human and animal health but presents significant challenges, particularly in grassroots health institutions. Our previous PDTCTR fluorescence sensing platform, which combined the engineered Cas12f1_ge4.1 system with recombinase polymerase amplification (RPA), significantly enhanced detection efficiency. However, its requirement for specialized equipment, costly RPA reagents, and absence of visual output restricted its practical application in such environments. To address these limitations, we developed the ERA/Cas12f1_ge4.1 system, integrating Cas12f1_ge4.1 with the cost-effective Enzymatic Recombinase Amplification (ERA). This system enables sensitive detection of Chlamydia psittaci double-stranded DNA within 50 min through both fluorescence and colloidal gold lateral flow assay strips. The platform achieves detection limits of 10 copies/μL for fluorescence and 100 copies/μL for lateral flow. Clinical validation involving 93 parrot samples demonstrated high performance in both detection modes. Fluorescence detection achieved 95.4 % sensitivity, 100 % specificity, a 100 % positive predictive value (PPV), and a 90.3 % negative predictive value (NPV). Meanwhile, the lateral flow assay exhibited 92.3 % sensitivity, 100 % specificity, 100 % PPV, and an 84.8 % NPV. The ERA/Cas12f1_ge4.1 system offers a rapid, accurate, cost-effective, and visually interpretable diagnostic tool suitable for both laboratory and community health centers. This advancement holds significant potential for improving psittacosis diagnosis and control, particularly in resource-limited environments.
鹦鹉热衣原体是鹦鹉热的病原体，快速准确地检测该病原体对人类和动物健康都至关重要，但面临重大挑战，尤其是在基层卫生机构。我们之前的PDTCTR荧光传感平台，将工程化的Cas12f1_ge4.1系统与重组酶聚合酶扩增（RPA）相结合，显著提高了检测效率。然而，其对专业设备的要求、昂贵的RPA试剂以及缺乏视觉输出限制了其在此类环境中的实际应用。为解决这些限制，我们开发了ERA/Cas12f1_ge4.1系统，将Cas12f1_ge4.1与经济高效的酶促重组扩增（ERA）相结合。该系统能够通过荧光和胶体金侧向流动检测试纸条在50分钟内灵敏检测鹦鹉热衣原体双链DNA。该平台荧光检测限为10拷贝/μL，侧向流动检测限为100拷贝/μL。涉及93份鹦鹉样本的临床验证表明，两种检测模式均具有高性能。荧光检测的灵敏度为95.4%，特异性为100%，阳性预测值（PPV）为100%，阴性预测值（NPV）为90.3%。同时，侧向流动检测的灵敏度为92.3%，特异性为100%，PPV为100%，NPV为84.8%。ERA/Cas12f1_ge4.1系统提供了一种快速、准确、经济高效且视觉上可解读的诊断工具，适用于实验室和社区卫生中心。这一进展对于改善鹦鹉热的诊断和控制具有巨大潜力，尤其是在资源有限的环境中。
