Rapid antimicrobial susceptibility testing (AST) based on in-situ time-lapse imaging of microcolonies
Globally, bacterial infections cause over seven million deaths annually, often attributable to delayed or inappropriate antibiotic administration. Antimicrobial susceptibility testing (AST) is routinely performed in clinical settings to guide antibiotic therapy; however, current methods are limited...
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| Veröffentlicht in: | Sensors and actuators. B, Chemical Jg. 448; S. 139012 |
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Elsevier B.V
01.02.2026
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| Abstract | Globally, bacterial infections cause over seven million deaths annually, often attributable to delayed or inappropriate antibiotic administration. Antimicrobial susceptibility testing (AST) is routinely performed in clinical settings to guide antibiotic therapy; however, current methods are limited by a minimum turnaround time of 16–20 h. Developing a rapid AST method based on bacterial morphological analysis could improve diagnostic efficiency and facilitate timely treatment for patients with severe bacterial infections. We developed an in-situ time-lapse imaging system for microcolonies (ISM-TLI). This system integrates an AST gel plate, a temperature-controlled incubation module capable of accommodating up to 30 patient samples simultaneously, a time-lapse imaging module, and integrated image processing algorithms. These algorithms perform image registration, microcolony identification, and minimum inhibitory concentration (MIC) interpretation. This system significantly reduces the turnaround time for obtaining AST profiles. A total of 21 antibiotics were tested on 9 reference strains, 24 clinical isolates and 2 urines samples, resulting in over 200 combinations. The concordance rates between the 2-hour and 3-hour TLI results and the broth microdilution (BMD) results were 90.8 % and 97.3 %, respectively. This system demonstrates the potential to shorten the existing clinical diagnostic process by over 14 h for isolates and 31–42 h for urine samples, significantly simplifying the process of guiding clinical antibiotic therapy.
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•AST within 3 h by in-situ time-lapse imaging of microcolonies on 96-well gel plate.•Fully automated system processes the incubation and imaging for up to 30 strains.•Accuracy > 97 % for 9 reference strains and 24 clinical isolates against 21 antibiotics.•Direct detection of AST in urine samples reduced the TAT from 1.5 days to 2–3 h. |
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| AbstractList | Globally, bacterial infections cause over seven million deaths annually, often attributable to delayed or inappropriate antibiotic administration. Antimicrobial susceptibility testing (AST) is routinely performed in clinical settings to guide antibiotic therapy; however, current methods are limited by a minimum turnaround time of 16–20 h. Developing a rapid AST method based on bacterial morphological analysis could improve diagnostic efficiency and facilitate timely treatment for patients with severe bacterial infections. We developed an in-situ time-lapse imaging system for microcolonies (ISM-TLI). This system integrates an AST gel plate, a temperature-controlled incubation module capable of accommodating up to 30 patient samples simultaneously, a time-lapse imaging module, and integrated image processing algorithms. These algorithms perform image registration, microcolony identification, and minimum inhibitory concentration (MIC) interpretation. This system significantly reduces the turnaround time for obtaining AST profiles. A total of 21 antibiotics were tested on 9 reference strains, 24 clinical isolates and 2 urines samples, resulting in over 200 combinations. The concordance rates between the 2-hour and 3-hour TLI results and the broth microdilution (BMD) results were 90.8 % and 97.3 %, respectively. This system demonstrates the potential to shorten the existing clinical diagnostic process by over 14 h for isolates and 31–42 h for urine samples, significantly simplifying the process of guiding clinical antibiotic therapy.
[Display omitted]
•AST within 3 h by in-situ time-lapse imaging of microcolonies on 96-well gel plate.•Fully automated system processes the incubation and imaging for up to 30 strains.•Accuracy > 97 % for 9 reference strains and 24 clinical isolates against 21 antibiotics.•Direct detection of AST in urine samples reduced the TAT from 1.5 days to 2–3 h. |
| ArticleNumber | 139012 |
| Author | Wu, Qingqing Song, Yizhi Wang, Ting Shen, Yuanheng Hu, Huijie Zhang, Changsong Li, Hui Zhang, Yingjun Xu, Yiwen Wang, Jingkai Meng, Siyu Zheng, Lesong Mao, Xin Zhang, Zhiwei Zhang, Zhiqiang Li, Jiang Wang, Yimai Zeng, Huan Tu, Junjie |
| Author_xml | – sequence: 1 givenname: Siyu surname: Meng fullname: Meng, Siyu organization: School of Biomedical Engineering (Suzhou), Division of Life Sciences and Medicine, University of Science and Technology of China, Suzhou, Jiangsu Province 215163, China – sequence: 2 givenname: Yiwen surname: Xu fullname: Xu, Yiwen organization: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, Jiangsu Province 215163, China – sequence: 3 givenname: Jingkai surname: Wang fullname: Wang, Jingkai organization: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, Jiangsu Province 215163, China – sequence: 4 givenname: Zhiwei surname: Zhang fullname: Zhang, Zhiwei organization: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, Jiangsu Province 215163, China – sequence: 5 givenname: Xin surname: Mao fullname: Mao, Xin organization: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, Jiangsu Province 215163, China – sequence: 6 givenname: Yimai surname: Wang fullname: Wang, Yimai organization: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, Jiangsu Province 215163, China – sequence: 7 givenname: Changsong surname: Zhang fullname: Zhang, Changsong organization: Department of Laboratory, The Affiliated Suzhou Hospital of Nanjing University Medical School (Suzhou Science and Technology Town Hospital), Suzhou, Jiangsu Province 215153, China – sequence: 8 givenname: Ting surname: Wang fullname: Wang, Ting organization: Department of Laboratory, The Affiliated Suzhou Hospital of Nanjing University Medical School (Suzhou Science and Technology Town Hospital), Suzhou, Jiangsu Province 215153, China – sequence: 9 givenname: Yuanheng surname: Shen fullname: Shen, Yuanheng organization: Department of Clinical Laboratory, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China – sequence: 10 givenname: Hui surname: Li fullname: Li, Hui organization: Hebei Institute of Drug and Medical Device Control, Shijiazhuang, Hebei Province 050000, China – sequence: 11 givenname: Lesong surname: Zheng fullname: Zheng, Lesong organization: School of Biomedical Engineering (Suzhou), Division of Life Sciences and Medicine, University of Science and Technology of China, Suzhou, Jiangsu Province 215163, China – sequence: 12 givenname: Qingqing surname: Wu fullname: Wu, Qingqing organization: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, Jiangsu Province 215163, China – sequence: 13 givenname: Huan surname: Zeng fullname: Zeng, Huan organization: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, Jiangsu Province 215163, China – sequence: 14 givenname: Junjie surname: Tu fullname: Tu, Junjie organization: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, Jiangsu Province 215163, China – sequence: 15 givenname: Yingjun surname: Zhang fullname: Zhang, Yingjun organization: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, Jiangsu Province 215163, China – sequence: 16 givenname: Zhiqiang surname: Zhang fullname: Zhang, Zhiqiang organization: School of Biomedical Engineering (Suzhou), Division of Life Sciences and Medicine, University of Science and Technology of China, Suzhou, Jiangsu Province 215163, China – sequence: 17 givenname: Huijie orcidid: 0009-0008-6112-7693 surname: Hu fullname: Hu, Huijie email: huhj@sibet.ac.cn organization: School of Biomedical Engineering (Suzhou), Division of Life Sciences and Medicine, University of Science and Technology of China, Suzhou, Jiangsu Province 215163, China – sequence: 18 givenname: Jiang surname: Li fullname: Li, Jiang email: 1013796363@qq.com organization: Laboratory Medicine Centre, Shenzhen Nanshan People's Hospital, Guangdong Province 518052, China – sequence: 19 givenname: Yizhi orcidid: 0000-0002-3005-382X surname: Song fullname: Song, Yizhi email: songyz@sibet.ac.cn organization: School of Biomedical Engineering (Suzhou), Division of Life Sciences and Medicine, University of Science and Technology of China, Suzhou, Jiangsu Province 215163, China |
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| Cites_doi | 10.3390/molecules24050892 10.1038/nprot.2007.521 10.3390/antibiotics8020052 10.1038/s41586-024-07725-1 10.1128/AAC.00979-20 10.1016/j.jinf.2017.08.012 10.1073/pnas.1819569116 10.34133/bmef.0017 10.1016/0168-1605(94)90165-1 10.1039/C6LC00417B 10.18520/cs/v125/i3/268-276 10.1039/C4LC01270D 10.1186/s12941-023-00644-5 10.3389/fmicb.2018.02888 10.1038/s41598-017-01278-2 10.1089/mdr.2017.0147 10.1086/647952 10.1098/rsif.2023.0730 10.1016/j.bios.2016.09.008 10.1039/D0SC01353F |
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Dis. doi: 10.1086/647952 – volume: 21 year: 2024 ident: 10.1016/j.snb.2025.139012_bib22 article-title: Multipad agarose plate: a rapid and high-throughput approach for antibiotic susceptibility testing publication-title: J. R. Soc. Interface doi: 10.1098/rsif.2023.0730 – year: 2023 ident: 10.1016/j.snb.2025.139012_bib20 – volume: 87 start-page: 669 year: 2017 ident: 10.1016/j.snb.2025.139012_bib11 article-title: A microfluidic device for antimicrobial susceptibility testing based on a broth dilution method publication-title: Biosens. Bioelectron. doi: 10.1016/j.bios.2016.09.008 – volume: 11 start-page: 6352 year: 2020 ident: 10.1016/j.snb.2025.139012_bib15 article-title: Microfluidic systems for rapid antibiotic susceptibility tests (ASTs) at the single-cell level publication-title: Chem. Sci. doi: 10.1039/D0SC01353F |
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| SubjectTerms | Antimicrobial susceptibility testing Image processing algorithm Microcolonies Time-lapse Imaging |
| Title | Rapid antimicrobial susceptibility testing (AST) based on in-situ time-lapse imaging of microcolonies |
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