State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, PR China; Guangdong Provincial Key Laboratory of Utilization and Conservation of Food and Medicinal Resources in Northern Region, Shaoguan University, Shaoguan, PR China; Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation, South China Agricultural University, Guangzhou 510642, PR China.
State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, PR China; Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation, South China Agricultural University, Guangzhou 510642, PR China.
Drug Resist Updat. 2024 Nov;77:101142. doi: 10.1016/j.drup.2024.101142. Epub 2024 Aug 24.
The spread of antibiotic resistance genes (ARGs), particularly those carried on plasmids, poses a major risk to global health. However, the extent and frequency of ARGs transfer in microbial communities among human, animal, and environmental sectors is not well understood due to a lack of effective tracking tools. We have developed a novel fluorescent tracing tool, CRISPR-AMRtracker, to study ARG transfer. It combines CRISPR/Cas9 fluorescence tagging, fluorescence-activated cell sorting, 16S rRNA gene sequencing, and microbial community analysis. CRISPR-AMRtracker integrates a fluorescent tag immediately downstream of ARGs, enabling the tracking of ARG transfer without compromising the host cell's antibiotic susceptibility, fitness, conjugation, and transposition. Notably, our experiments demonstrate that sfGFP-tagged plasmid-borne mcr-1 can transfer across diverse bacterial species within fecal samples. This innovative approach holds the potential to illuminate the dynamics of ARG dissemination and provide valuable insights to shape effective strategies in mitigating the escalating threat of antibiotic resistance.
抗生素耐药基因（ARGs）的传播，特别是那些携带在质粒上的基因，对全球健康构成了重大威胁。然而，由于缺乏有效的跟踪工具，人类、动物和环境领域中微生物群落中 ARGs 转移的程度和频率还不是很清楚。我们开发了一种新的荧光跟踪工具 CRISPR-AMRtracker，用于研究 ARG 转移。它结合了 CRISPR/Cas9 荧光标记、荧光激活细胞分选、16S rRNA 基因测序和微生物群落分析。CRISPR-AMRtracker 将荧光标签直接整合到 ARGs 的下游，在不影响宿主细胞抗生素敏感性、适应性、接合和转座的情况下，能够跟踪 ARG 转移。值得注意的是，我们的实验表明，sfGFP 标记的质粒携带的 mcr-1 可以在粪便样本中的不同细菌物种之间转移。这种创新方法有可能阐明 ARG 传播的动态，并为制定有效的策略提供有价值的见解，以减轻抗生素耐药性不断升级的威胁。
