Zhou Zihan, Liang Lina, Liao Chuan, Pan Lele, Wang Chunfang, Ma Jiangmei, Yi Xueli, Tan Meiying, Li Xuebin, Wei Guijiang
Center for Medical Laboratory Science, Affiliated Hospital of Youjiang Medical University for Nationalities, Baise, Guangxi, China.
Baise Key Laboratory for Research and Development on Clinical Molecular Diagnosis for High-Incidence Diseases, Baise, Guangxi, China.
Front Microbiol. 2024 Mar 6;15:1359976. doi: 10.3389/fmicb.2024.1359976. eCollection 2024.
Carbapenem-resistant (CRAB) poses a severe nosocomial threat, prompting a need for efficient detection methods. Traditional approaches, such as bacterial culture and PCR, are time-consuming and cumbersome. The CRISPR-based gene editing system offered a potential approach for point-of-care testing of CRAB.
We integrated recombinase polymerase amplification (RPA) and CRISPR-Cas12a system to swiftly diagnose CRAB-associated genes, and . This multiplex RPA-CRISPR-Cas12a system eliminates bulky instruments, ensuring a simplified UV lamp-based outcome interpretation.
Operating at 37°C to 40°C, the entire process achieves CRAB diagnosis within 90 minutes. Detection limits for and genes are 1.3 × 10 ng/μL, exhibiting exclusive CRAB detection without cross-reactivity to common pathogens. Notably, the platform shows 100% concordance with PCR when testing 30 clinical strains.
In conclusion, our multiplex RPA coupled with the CRISPR-Cas12a system provides a fast and sensitive CRAB detection method, overcoming limitations of traditional approaches and holding promise for efficient point-of-care testing.
耐碳青霉烯类鲍曼不动杆菌（CRAB）构成严重的医院感染威胁，因此需要高效的检测方法。传统方法，如细菌培养和聚合酶链反应（PCR），耗时且繁琐。基于CRISPR的基因编辑系统为CRAB的即时检测提供了一种潜在方法。
我们整合了重组酶聚合酶扩增（RPA）和CRISPR-Cas12a系统，以快速诊断与CRAB相关的基因。这种多重RPA-CRISPR-Cas12a系统无需大型仪器，确保基于紫外线灯的结果解读得以简化。
该系统在37°C至40°C下运行，整个过程在90分钟内即可完成CRAB诊断。和基因的检测限为1.3×10 ng/μL，能特异性检测CRAB，对常见病原体无交叉反应。值得注意的是，在检测30株临床菌株时，该平台与PCR的结果一致性达100%。
总之，我们的多重RPA与CRISPR-Cas12a系统相结合，提供了一种快速且灵敏的CRAB检测方法，克服了传统方法的局限性，有望实现高效的即时检测。
