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ORIGINAL RESEARCH article
Front. Cell. Infect. Microbiol.
Sec. Clinical Infectious Diseases
Volume 14 - 2024 |
doi: 10.3389/fcimb.2024.1409078
An accurate and convenient method for Mycoplasma pneumoniae via one-step LAMP-CRISPR/Cas12b detection platform
Provisionally accepted
Tao Liu
1
Qing Liu
2,3
Fuqun Chen
1*
Ying Shi
1*
Guliya •. Maimaiti
1*
Zhanhua Yang
1*
Shutao Zheng
3,4*
Xiaomei LU
3,4
Hui Li
1*
Zhaoyun Chen
1*- 1 Department of Clinical Laboratory, First Affiliated Hospital of Xinjiang Medical University, Urumqi, China
- 2 First Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang Uyghur Region, China
- 3 Key Laboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases, Xinjiang Medical University, Urumqi, Xinjiang, China
- 4 Clinical Medical College, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang, China
Mycoplasma pneumoniae (MP) is the major cause of respiratory infections that threaten the health of children and adolescents worldwide. Therefore, an early, simple, and accurate detection approach for MP is critical to prevent outbreaks of MP-induced community-acquired pneumonia. Here, we explored a simple and accurate method for MP identification that combines loop-mediated isothermal amplification (LAMP) with the CRISPR/Cas12b assay in a one-pot reaction. In the current study, the whole reaction was completed within 1 h at a constant temperature of 57℃. The limit of detection of this assay was 33.7 copies per reaction. The specificity of the LAMP-CRISPR/Cas12b method was 100%, without any cross-reactivity with other pathogens. Overall, 183 clinical samples were used to evaluate the clinical performance of LAMP-CRISPR/Cas12b. Compared with the gold standard results from real-time PCR, the present method provided a sensitivity of 88.11% (126/143), specificity of 100% (129/129), and consistency of 93.75% (255/272). Taken together, our preliminary results illustrate that the LAMP-CRISPR/Cas12b method is a simple and reliable tool for MP diagnosis that can be performed in resource-limited regions.
Keywords: Mycoplasma pneumoniae, LAMP, CRISPR, Cas12b, One-step, One-pot
Received: 29 Mar 2024; Accepted: 22 Jul 2024.
Copyright: © 2024 Liu, Liu, Chen, Shi, Maimaiti, Yang, Zheng, LU, Li and Chen. 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:
Fuqun Chen, Department of Clinical Laboratory, First Affiliated Hospital of Xinjiang Medical University, Urumqi, China
Ying Shi, Department of Clinical Laboratory, First Affiliated Hospital of Xinjiang Medical University, Urumqi, China
Guliya •. Maimaiti, Department of Clinical Laboratory, First Affiliated Hospital of Xinjiang Medical University, Urumqi, China
Zhanhua Yang, Department of Clinical Laboratory, First Affiliated Hospital of Xinjiang Medical University, Urumqi, China
Shutao Zheng, Clinical Medical College, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, 830054, Xinjiang, China
Hui Li, Department of Clinical Laboratory, First Affiliated Hospital of Xinjiang Medical University, Urumqi, China
Zhaoyun Chen, Department of Clinical Laboratory, First Affiliated Hospital of Xinjiang Medical University, Urumqi, China
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