ORIGINAL RESEARCH article
Front. Bioeng. Biotechnol.
Sec. Biosensors and Biomolecular Electronics
Volume 13 - 2025 | doi: 10.3389/fbioe.2025.1663915
A light-controlled one-tube detection platform combining CRISPR-Cas12a and RPA: An innovative approach for rapid diagnosis of Acinetobacter baumannii
Provisionally accepted
- Youjiang Medical University for Nationalities, Baise, China
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Acinetobacter baumannii (A. baumannii) is a significant pathogen associated with nosocomial infections, predominantly affecting immunocompromised patients, and is linked to high mortality rates. To control infection rates, there is an urgent need for a diagnostic method that is cost-effective, rapid, and user-friendly, meeting the current demand for timely diagnosis. We have developed a one-tube detection method for UV light unlocking based on RPA-CRISPR/Cas12a technology. This method utilizes the photodegradable chemical group NPOM-dt to chemically modify the crRNA base, preventing it from complementary pairing with the base of the target molecule, thereby temporarily silencing the CRISPR system. After RPA preamplification, the caged modification group on the crRNA was removed with brief irradiation with ultraviolet light to restore the activity of the CRISPR/Cas12a system. This light-controlled one-tube system simplifies operation while significantly reducing the risk of contamination caused by repeated tube openings. Our results demonstrated that the detection platform achieved a limit of detection as low as 10 copies/ÎĽL for target fragments, with no cross-reactivity observed with genomic DNA from six clinically common pathogenic bacteria, showcasing excellent sensitivity and specificity.Additionally, clinical validation was performed using 38 sputum samples. The system successfully identified A. baumannii in sputum specimens, with results consistent with those obtained via conventional PCR. These findings highlight the potential of the light-controlled one-tube RPA-CRISPR/Cas12a system as a promising diagnostic tool.
Keywords: CRISPR/Cas12a, NPOM-dt, One-tube detection, POCT, Acinetobacter baumannii
Received: 11 Jul 2025; Accepted: 29 Jul 2025.
Copyright: © 2025 Liang, Zhou, Pan, Luo, Ma, Ren, Li and Wei. 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:
Lina Liang, Youjiang Medical University for Nationalities, Baise, China
Xuebin Li, Youjiang Medical University for Nationalities, Baise, China
Guijiang Wei, Youjiang Medical University for Nationalities, Baise, China
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