Rapid genotypic antibiotic susceptibility test using CRISPR-Cas12a for urinary tract infection

• Published in: Analyst (London) Vol. 145; no. 15; pp. 5226 - 5231
• Main Authors: Chen, Juhong, Jiang, Fuguo, Huang, Chao-Wei, Lin, Liwei
• Format: Journal Article
• Language: English
• Published: England Royal Society of Chemistry 07.08.2020
• Subjects: Amplification; Anti-Bacterial Agents - pharmacology; Antibiotics; Bacteria; Bacteria - genetics; Clustered Regularly Interspaced Short Palindromic Repeats; CRISPR; CRISPR-Cas Systems - genetics; Humans; Infectious diseases; Urinary tract; Urinary Tract Infections - diagnosis; Urinary Tract Infections - drug therapy; Urogenital system
• ISSN: 0003-2654; 1364-5528
• DOI: 10.1039/d0an00947d

The current clinical protocol to conduct a bacterial antibiotic susceptibility test (AST) requires at least 18 hours, and cannot be accomplished during a single visit for patients. Here, a new method based on the technique of CRISPR-Cas12a is utilized to accomplish a bacterial genotypic AST within one hour with good accuracy. Two amplification approaches are employed and compared: (1) enriching the bacterial concentration by culturing in growth media; and (2) amplifying target DNA from raw samples by recombinase polymerase amplification (RPA). The results show that CRISPR combined with RPA can rapidly and accurately provide a bacterial genotypic AST of urine samples with urinary tract infections for precise antibiotic treatment. As such, this technology could open a new class of rapid bacterial genotypic AST for various infectious diseases. A new method based on the technique of CRISPR-Cas12a is developed to accomplish a bacterial antibiotic susceptibility test (AST) within one hour with good accuracy.