Exploiting β‐Lactams‐Induced Lysis and DNA Fragmentation for Rapid Molecular Antimicrobial Susceptibility Testing of Neisseria Gonorrhoeae via Dual‐Digital PCR
Abstract 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 discoverin...
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Wiley
2024-12-01
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| Series: | Advanced Science |
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| Online Access: | https://doi.org/10.1002/advs.202405272 |
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| author | Jiumei Hu Liben Chen Pengfei Zhang Fan‐En Chen Hui Li Kuangwen Hsieh Sixuan Li Johan H. Melendez Tza‐Huei Wang |
| author_facet | Jiumei Hu Liben Chen Pengfei Zhang Fan‐En Chen Hui Li Kuangwen Hsieh Sixuan Li Johan H. Melendez Tza‐Huei Wang |
| author_sort | Jiumei Hu |
| collection | DOAJ |
| description | Abstract 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. |
| format | Article |
| id | doaj-art-ebe74f555ca24706b83e02d6a6590f3e |
| institution | DOAJ |
| issn | 2198-3844 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Wiley |
| record_format | Article |
| series | Advanced Science |
| spelling | doaj-art-ebe74f555ca24706b83e02d6a6590f3e2025-08-20T02:50:00ZengWileyAdvanced Science2198-38442024-12-011146n/an/a10.1002/advs.202405272Exploiting β‐Lactams‐Induced Lysis and DNA Fragmentation for Rapid Molecular Antimicrobial Susceptibility Testing of Neisseria Gonorrhoeae via Dual‐Digital PCRJiumei Hu0Liben Chen1Pengfei Zhang2Fan‐En Chen3Hui Li4Kuangwen Hsieh5Sixuan Li6Johan H. Melendez7Tza‐Huei Wang8Department of Mechanical Engineering Johns Hopkins University Baltimore MD 21218 USADepartment of Mechanical Engineering Johns Hopkins University Baltimore MD 21218 USADepartment of Biomedical Engineering Johns Hopkins School of Medicine Baltimore MD 21205 USADepartment of Biomedical Engineering Johns Hopkins School of Medicine Baltimore MD 21205 USADepartment of Mechanical Engineering Johns Hopkins University Baltimore MD 21218 USADepartment of Mechanical Engineering Johns Hopkins University Baltimore MD 21218 USADepartment of Mechanical Engineering Johns Hopkins University Baltimore MD 21218 USADivision of Infectious Diseases Department of Medicine Johns Hopkins University School of Medicine Baltimore MD 21205 USADepartment of Mechanical Engineering Johns Hopkins University Baltimore MD 21218 USAAbstract 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.https://doi.org/10.1002/advs.202405272antimicrobial resistancebacterial lysisDNA fragmentationmicrofluidic dual‐digital PCRNesseria gonorrhoeae (N. gonorrheoae) |
| spellingShingle | Jiumei Hu Liben Chen Pengfei Zhang Fan‐En Chen Hui Li Kuangwen Hsieh Sixuan Li Johan H. Melendez Tza‐Huei Wang Exploiting β‐Lactams‐Induced Lysis and DNA Fragmentation for Rapid Molecular Antimicrobial Susceptibility Testing of Neisseria Gonorrhoeae via Dual‐Digital PCR Advanced Science antimicrobial resistance bacterial lysis DNA fragmentation microfluidic dual‐digital PCR Nesseria gonorrhoeae (N. gonorrheoae) |
| title | Exploiting β‐Lactams‐Induced Lysis and DNA Fragmentation for Rapid Molecular Antimicrobial Susceptibility Testing of Neisseria Gonorrhoeae via Dual‐Digital PCR |
| title_full | Exploiting β‐Lactams‐Induced Lysis and DNA Fragmentation for Rapid Molecular Antimicrobial Susceptibility Testing of Neisseria Gonorrhoeae via Dual‐Digital PCR |
| title_fullStr | Exploiting β‐Lactams‐Induced Lysis and DNA Fragmentation for Rapid Molecular Antimicrobial Susceptibility Testing of Neisseria Gonorrhoeae via Dual‐Digital PCR |
| title_full_unstemmed | Exploiting β‐Lactams‐Induced Lysis and DNA Fragmentation for Rapid Molecular Antimicrobial Susceptibility Testing of Neisseria Gonorrhoeae via Dual‐Digital PCR |
| title_short | Exploiting β‐Lactams‐Induced Lysis and DNA Fragmentation for Rapid Molecular Antimicrobial Susceptibility Testing of Neisseria Gonorrhoeae via Dual‐Digital PCR |
| title_sort | exploiting β lactams induced lysis and dna fragmentation for rapid molecular antimicrobial susceptibility testing of neisseria gonorrhoeae via dual digital pcr |
| topic | antimicrobial resistance bacterial lysis DNA fragmentation microfluidic dual‐digital PCR Nesseria gonorrhoeae (N. gonorrheoae) |
| url | https://doi.org/10.1002/advs.202405272 |
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