Cas14VIDet: A visual instant method free from PAM restriction for antibiotic resistance bacteria detection.

Autor: Lai Y; Department of Gastroenterology, Ganzhou People's Hospital Affiliated to Nanchang University, Ganzhou, 341000, China; Department of Gastroenterology, Shanghai Changhai Hospital, Naval Medical University, Shanghai, 200433, China. Electronic address: xhyongkanglai@hotmail.com., Guo K; Human Phenome Institute, State Key Laboratory of Genetic Engineering, Fudan University, Shanghai, 200438, China., Zhu C; Department of Gastroenterology, Ganzhou People's Hospital Affiliated to Nanchang University, Ganzhou, 341000, China; Department of Gastroenterology, Shanghai Changhai Hospital, Naval Medical University, Shanghai, 200433, China., Lv Y; Department of Gastroenterology, Shanghai Changhai Hospital, Naval Medical University, Shanghai, 200433, China., Wu H; Department of Orthopaedic Surgery, The Affiliated Hospital of Qingdao University, No. 16 Jiangsu Road, Qingdao, 266000, China., Zhang L; Department of Gastroenterology, Shanghai Changhai Hospital, Naval Medical University, Shanghai, 200433, China., Jiang M; Department of Gastroenterology, Shanghai Changhai Hospital, Naval Medical University, Shanghai, 200433, China., Pang Y; Department of Gastroenterology, Shanghai Changhai Hospital, Naval Medical University, Shanghai, 200433, China., Zhao J; Department of Gastroenterology, Shanghai Changhai Hospital, Naval Medical University, Shanghai, 200433, China. Electronic address: jlzhao9@163.com., Wang R; Human Phenome Institute, State Key Laboratory of Genetic Engineering, Fudan University, Shanghai, 200438, China; Center for Medical Research and Innovation, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai, 201399, China; International Human Phenome Institutes, Shanghai, 200438, China. Electronic address: wangr@fudan.edu.cn., Li Z; Department of Gastroenterology, Ganzhou People's Hospital Affiliated to Nanchang University, Ganzhou, 341000, China; Department of Gastroenterology, Shanghai Changhai Hospital, Naval Medical University, Shanghai, 200433, China. Electronic address: zhsl@vip.163.com.
Jazyk: angličtina
Zdroj: Biosensors & bioelectronics [Biosens Bioelectron] 2025 Jan 15; Vol. 268, pp. 116884. Date of Electronic Publication: 2024 Nov 08.
DOI: 10.1016/j.bios.2024.116884
Abstrakt: A personalized treatment strategy that selects sensitive antibiotics based on Helicobacter pylori (H. pylori) resistance genes is currently the most effective approach to address the challenge of H. pylori eradication. However, the widespread adoption of this strategy is hindered by the long processing times and high costs associated with traditional resistance gene detection methods. In this study, we combined ultra-fast PCR with CRISPR/Cas14 into a single reaction system, establishing a rapid, one-pot visual platform named Cas14VIDet (Cas14-based Visual Instant Detection) for detecting H. pylori resistance genes. Cas14VIDet does not require a PAM sequence and excels in identifying single nucleotide polymorphisms, with the detection sensitivity approaching the level of a single bacterial colony (10 0  CFU/mL). The entire detection process can be completed within 10 min, and results are directly observable with the naked eye. We validated Cas14VIDet by testing 50 clinical samples and compared it with Sanger sequencing. The results showed that Cas14VIDet achieved 100% sensitivity, 100% specificity, and 100% accuracy in detecting H. pylori resistance genes to levofloxacin. Therefore, we believe this method holds great potential for rapid detection of H. pylori resistance, potentially supporting personalized treatment of H. pylori infections in the future.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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