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ORIGINAL RESEARCH article
Front. Pediatr.
Sec. Pediatric Infectious Diseases
Volume 13 - 2025 | doi: 10.3389/fped.2025.1533100
Method Development and Clinical Validation of LAMP-CRISPR/Cas12a for Rapid Detection of Respiratory Pathogens in Children
Provisionally accepted- Bethune First Hospital of Jilin University, Changchun, China
Background: Respiratory tract infections pose a substantial health burden, particularly among pediatric populations globally. The timely and accurate identification of pathogens such as Streptococcus pneumoniae (SP) and Mycoplasma pneumoniae (MP) is critical for effective clinical management.Methods: In this study, a novel diagnostic approach combining loop-mediated isothermal amplification (LAMP) with CRISPR-Cas12a technology was developed for detecting SP and MP in clinical respiratory samples. A total of 23 specimens, including bronchoalveolar lavage fluid and nasopharyngeal swab samples, were assessed to evaluate the feasibility and performance of the method. After nucleic acid extraction, samples underwent LAMP amplification followed by CRISPR-Cas12a-mediated fluorescence detection.Results: The LAMP-CRISPR/Cas12a method demonstrated high sensitivity and specificity for SP detection. It exhibited excellent sensitivity for SP and promising specificity for MP. Comparative analysis with standard diagnostic methods highlighted its potential to enhance diagnostic accuracy and efficiency. The assay provided results within 1 h, which is suitable for rapid point-of-care testing.Conclusion: The integrated LAMP-CRISPR/Cas12a approach represents a significant advancement in detecting respiratory pathogens in clinical settings. It offers a rapid, sensitive, and specific diagnostic tool for identifying SP and MP, which is crucial for guiding precision therapies and improving patient outcomes. Future research aims to optimize assay sensitivity, streamline workflow to minimize contamination risks, and expand its detection scope so that other types of pathogens and mutation resistance genes can be detected. This molecular diagnostic strategy holds promise for the management of respiratory infections by enabling early and precise pathogen identification.
Keywords: Respiratory Tract Infections, LAMP-CRISPR/Cas12a, Streptococcus pneumoniae, Mycoplasma pneumoniae, Molecular diagnostics
Received: 23 Nov 2024; Accepted: 20 Mar 2025.
Copyright: © 2025 Zhou, Zhao and Meng. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Fanzheng Meng, Bethune First Hospital of Jilin University, Changchun, China
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