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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\u2010cell techniques, including live\u2010cell imaging, flow cytometry, and digital polymerase chain reaction (PCR) are utilized, to investigate the lysis dynamics and molecular features of <jats:italic>N. gonorrhoeae<\/jats:italic> upon exposure to \u03b2\u2010lactam antimicrobials. Distinct patterns of bacterial lysis and DNA fragmentation are uncovered in susceptible strains. Leveraging these discoveries, a microfluidic dual\u2010digital PCR approach that combines single\u2010cell and single\u2010molecule analyses, facilitate rapid and efficient phenotypic molecular AST for <jats:italic>N. gonorrhoeae<\/jats:italic> against \u03b2\u2010lactams is developed. This proof\u2010of\u2010concept 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.<\/jats:p>","DOI":"10.1002\/advs.202405272","type":"journal-article","created":{"date-parts":[[2024,10,18]],"date-time":"2024-10-18T06:39:29Z","timestamp":1729233569000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Exploiting \u03b2\u2010Lactams\u2010Induced Lysis and DNA Fragmentation for Rapid Molecular Antimicrobial Susceptibility Testing of <i>Neisseria Gonorrhoeae<\/i> via Dual\u2010Digital PCR"],"prefix":"10.1002","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9774-9090","authenticated-orcid":false,"given":"Jiumei","family":"Hu","sequence":"first","affiliation":[{"name":"Department of Mechanical Engineering Johns Hopkins University  Baltimore MD 21218 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Liben","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering Johns Hopkins University  Baltimore MD 21218 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pengfei","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering Johns Hopkins School of Medicine  Baltimore MD 21205 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fan\u2010En","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering Johns Hopkins School of Medicine  Baltimore MD 21205 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hui","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering Johns Hopkins University  Baltimore MD 21218 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kuangwen","family":"Hsieh","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering Johns Hopkins University  Baltimore MD 21218 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sixuan","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering Johns Hopkins University  Baltimore MD 21218 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Johan H.","family":"Melendez","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases Department of Medicine Johns Hopkins University School of Medicine  Baltimore MD 21205 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3540-9354","authenticated-orcid":false,"given":"Tza\u2010Huei","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering Johns Hopkins University  Baltimore MD 21218 USA"},{"name":"Department of Biomedical Engineering Johns Hopkins School of Medicine  Baltimore MD 21205 USA"},{"name":"Institute for NanoBiotechnology Johns Hopkins University  Baltimore MD 21218 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,10,18]]},"reference":[{"key":"e_1_2_8_1_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(21)02724-0"},{"key":"e_1_2_8_2_1","doi-asserted-by":"crossref","unstructured":"B. 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