The Institute of Pathogenic Biology, Hengyang Medical School, University of South China, Hengyang, China.
Department of Clinical Laboratory, The Second People's Hospital of Foshan, Foshan, China.
Front Cell Infect Microbiol. 2024 Nov 1;14:1477422. doi: 10.3389/fcimb.2024.1477422. eCollection 2024.
is a specialized intracellular parasitic pathogen capable of causing pneumonia, sinusitis, bronchitis, and other respiratory diseases, which pose significant public health challenges. Therefore, rapid, accurate, and sensitive diagnosis is crucial for the prevention and treatment of respiratory diseases caused by . In this study, we combined enzymatic recombination amplification (ERA) with the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) 12a system (CRISPR/Cas12a) to develop a dual detection platform termed the Cpn-ERA-CRISPR/Cas12a dual system. This system integrates both the ERA-CRISPR/Cas12a fluorescence system and the ERA-CRISPR/Cas12a lateral flow system. Detection results can be measured using a fluorescence detector or observed with the naked eye on lateral flow strips. The fluorescence system and the lateral flow system detect in 30 minutes and 15 minutes, respectively. This dual system exhibits no cross-reactivity with the other seven pathogens, demonstrating high specificity, and achieves a sensitivity of 10 copies/µL. Additionally, the Cpn-ERA-CRISPR/Cas12a dual system was employed to analyze 39 clinical samples, comprising 19 positive and 20 negative samples. The detection rate for positive samples was 100%, with no positive results in the negative samples, indicating a high level of concordance with qPCR results. In summary, the Cpn-ERA-CRISPR/Cas12a dual system represents a novel tool for diagnosing and holds promising application potential in grassroots community hospitals.
是一种专门的细胞内寄生病原体，能够引起肺炎、鼻窦炎、支气管炎和其他呼吸道疾病，对公共卫生构成重大挑战。因此，快速、准确、敏感的诊断对于预防和治疗 引起的呼吸道疾病至关重要。在本研究中，我们将酶促重组扩增（ERA）与成簇规律间隔短回文重复（CRISPR）/CRISPR 相关蛋白（Cas）12a 系统（CRISPR/Cas12a）相结合，开发了一种称为 Cpn-ERA-CRISPR/Cas12a 双检测平台。该系统集成了 ERA-CRISPR/Cas12a 荧光系统和 ERA-CRISPR/Cas12a 侧流系统。可以使用荧光检测器测量检测结果，也可以在侧流条上用肉眼观察。荧光系统和侧流系统分别在 30 分钟和 15 分钟内检测到 。该双系统与其他七种病原体无交叉反应，表现出高度特异性，灵敏度达到 10 拷贝/μL。此外，Cpn-ERA-CRISPR/Cas12a 双系统用于分析 39 个临床样本，包括 19 个阳性样本和 20 个阴性样本。阳性样本的检测率为 100%，阴性样本无阳性结果，与 qPCR 结果高度一致。综上所述，Cpn-ERA-CRISPR/Cas12a 双系统为 诊断提供了一种新工具，在基层社区医院具有广阔的应用潜力。
