Hu Jiumei, Chen Liben, Zhang Pengfei, Chen Fan-En, Li Hui, Hsieh Kuangwen, Li Sixuan, Melendez Johan H, Wang Tza-Huei
Department of Mechanical Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA.
Department of Biomedical Engineering, Johns Hopkins School of Medicine, Baltimore, MD, 21205, USA.
Adv Sci (Weinh). 2024 Dec;11(46):e2405272. doi: 10.1002/advs.202405272. Epub 2024 Oct 18.
The evolution of antimicrobial resistance (AMR) presents substantial challenges to global medical health systems. Neisseria gonorrhoeae (N. gonorrhoeae), in particular, has developed resistance to all currently available antimicrobials. Addressing this issue necessitates not only discovering new antimicrobials but also deepening the understanding of bacterial responses to these agents, which can lead to new markers for rapid antimicrobial susceptibility testing (AST). Such advancements can enhance treatment outcomes and promote antimicrobial stewardship. In this study, single-cell techniques, including live-cell imaging, flow cytometry, and digital polymerase chain reaction (PCR) are utilized, to investigate the lysis dynamics and molecular features of N. gonorrhoeae upon exposure to β-lactam antimicrobials. Distinct patterns of bacterial lysis and DNA fragmentation are uncovered in susceptible strains. Leveraging these discoveries, a microfluidic dual-digital PCR approach that combines single-cell and single-molecule analyses, facilitate rapid and efficient phenotypic molecular AST for N. gonorrhoeae against β-lactams is developed. This proof-of-concept validation demonstrates the effectiveness of the method in accessing antimicrobial susceptibility across a range of bacterial strains, contributing valuable insights for advancing the battle against AMR.
抗菌药物耐药性（AMR）的演变给全球医疗卫生系统带来了巨大挑战。尤其是淋病奈瑟菌（N. gonorrhoeae），已对目前所有可用的抗菌药物产生了耐药性。解决这一问题不仅需要发现新的抗菌药物，还需要加深对细菌对这些药物反应的理解，这可能会带来用于快速抗菌药物敏感性测试（AST）的新标志物。这些进展可以改善治疗效果并促进抗菌药物管理。在本研究中，利用包括活细胞成像、流式细胞术和数字聚合酶链反应（PCR）在内的单细胞技术，研究淋病奈瑟菌在接触β-内酰胺类抗菌药物后的裂解动力学和分子特征。在敏感菌株中发现了不同的细菌裂解和DNA片段化模式。利用这些发现，开发了一种结合单细胞和单分子分析的微流控双数字PCR方法，用于快速、高效地对淋病奈瑟菌针对β-内酰胺类药物进行表型分子AST。这种概念验证验证证明了该方法在评估一系列细菌菌株抗菌药物敏感性方面的有效性，为推进对抗AMR的斗争提供了有价值的见解。
