Department of Biomaterials, The Higher Educational Key Laboratory for Biomedical Engineering of Fujian Province, Research Center of Biomedical Engineering of Xiamen, College of Materials, Xiamen University, Xiamen, 361005, PR China.
School of Pharmaceutical Sciences, Xiamen University, Xiamen, 361102, PR China.
Anal Chim Acta. 2023 Apr 22;1251:341014. doi: 10.1016/j.aca.2023.341014. Epub 2023 Feb 25.
The accumulation and spread of antibiotic resistance bacteria (ARB) in the environment may accelerate the formation of superbugs and seriously threaten the health of all living beings. The timeliness and accurate diagnosing of antibiotic resistance is essential to controlling the propagation of superbugs in the environment and formulating effective public health management programs. Herein, we developed a speedy, sensitive, accurate, and user-friendly colorimetric assay for antibiotic resistance, via a synergistic combination of the peroxidase-like property of the Au-FeO nanozyme and the specific gene identification capability of the CRISPR-Cas12a. Once the CRISPR-Cas12a system recognizes a target resistance gene, it activates its trans-cleavage activity and subsequently releases the Au-FeO nanozymes, which oxidizes the 3,3,5,5-tetramethylbenzidine (TMB) with color change from transparent to blue. The diagnosing signals could be captured and analyzed by a smartphone. This method detected kanamycin-resistance genes, ampicillin-resistance genes, and chloramphenicol-resistance genes by simple operation steps with high sensitivity (<0.1 CFU μL) and speediness (<1 h). This approach may prove easy for the accurate and sensitive diagnosis of the ARGs or ARB in the field, thus surveilling and controlling the microbial water quality flexibly and efficiently.
抗生素耐药菌（ARB）在环境中的积累和传播可能会加速超级细菌的形成，严重威胁所有生物的健康。及时准确地诊断抗生素耐药性对于控制环境中超细菌的传播和制定有效的公共卫生管理计划至关重要。在此，我们通过金-铁氧纳米酶的过氧化物酶样特性和 CRISPR-Cas12a 的特定基因识别能力的协同组合，开发了一种快速、敏感、准确且易于使用的抗生素耐药性比色分析方法。一旦 CRISPR-Cas12a 系统识别出目标耐药基因，它就会激活其转录切割活性，随后释放金-铁氧纳米酶，纳米酶将 3,3,5,5-四甲基联苯胺（TMB）氧化，使颜色从透明变为蓝色。通过智能手机可以捕获和分析诊断信号。该方法通过简单的操作步骤检测卡那霉素耐药基因、氨苄西林耐药基因和氯霉素耐药基因，具有高灵敏度（<0.1 CFU μL）和快速（<1 小时）的特点。该方法可用于现场准确、灵敏地检测 ARGs 或 ARB，从而灵活、高效地监测和控制微生物水质。
