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<p>Antibiotic resistance poses a significant global public health challenge, particularly concerning carbapenem-resistant <i>Klebsiella pneumoniae</i> (CRKP). A Raman-based antimicrobial susceptibility testing (R-AST) method was developed, utilizing D₂O-labeled single-cell Raman spectroscopy alongside the Metabolic Inhibition Level (MIL) for rapid CRKP antimicrobial susceptibility testing and carbapenemase enzyme typing. By analyzing 30,000 single-cell Raman spectra from 57 <i>K. pneumoniae</i> strains, the R-AST accurately detected CRKP within two hours. Furthermore, R-AST was able to differentiate between amikacin-sensitive and amikacin-resistant strains within the CRKP category, thereby providing potential treatment options. Notably, both R-AST classifications demonstrated 100% category agreement with the Vitek<sup>®</sup>-2 Compact system. The R-AST demonstrated greater accuracy in distinguishing carbapenemase types compared to the carbapenemase inhibitor enhancement test in <i>K. pneumoniae</i>, achieving an overall accuracy of 95.74% (95% CI: 85.46–98.77%) in enzyme typing. However, it misclassified 2 out of 21 KPC-producing CRKP strains as KPC and NDM-coproducing CRKP strains. Conversely, the overall accuracy of the carbapenemase inhibitor enhancement test was 93.62% (95% CI: 82.69–97.66%), with 3 out of 9 KPC and NDM-coproducing strains being missed. By delivering reliable results within 2&#xa0;h, the R-AST provides an efficient, dependable, and broadly applicable approach to rapid antimicrobial susceptibility testing in clinical settings, potentially mitigating antibiotic misuse and the spread of drug-resistant bacteria.</p> Graphical Abstract <p></p>

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Accelerated detection of carbapenem resistance in Klebsiella pneumoniae via single-cell Raman spectroscopy

  • 期刊论文

  • 发表日期：

    2025年4月29日

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  • Jia-Cheng Ding,
  • Li-Ting Dai,
  • Jia-Xin Tang,
  • Hui-Min Zhang,
  • Le-Le Zhai,
  • Chuang-Ye Cai,
  • Zhi-Rou Xiao,
  • Zhe-Mei Chen,
  • Jia-Hui Ling,
  • Meng-Xue Zou,
  • Xi-Wu Cao,
  • Li-Juan Lin,
  • Zi-Heng Xu,
  • Pei-Bo Yuan,
  • Ding-Qiang Chen

摘要

Antibiotic resistance poses a significant global public health challenge, particularly concerning carbapenem-resistant Klebsiella pneumoniae (CRKP). A Raman-based antimicrobial susceptibility testing (R-AST) method was developed, utilizing D₂O-labeled single-cell Raman spectroscopy alongside the Metabolic Inhibition Level (MIL) for rapid CRKP antimicrobial susceptibility testing and carbapenemase enzyme typing. By analyzing 30,000 single-cell Raman spectra from 57 K. pneumoniae strains, the R-AST accurately detected CRKP within two hours. Furthermore, R-AST was able to differentiate between amikacin-sensitive and amikacin-resistant strains within the CRKP category, thereby providing potential treatment options. Notably, both R-AST classifications demonstrated 100% category agreement with the Vitek®-2 Compact system. The R-AST demonstrated greater accuracy in distinguishing carbapenemase types compared to the carbapenemase inhibitor enhancement test in K. pneumoniae, achieving an overall accuracy of 95.74% (95% CI: 85.46–98.77%) in enzyme typing. However, it misclassified 2 out of 21 KPC-producing CRKP strains as KPC and NDM-coproducing CRKP strains. Conversely, the overall accuracy of the carbapenemase inhibitor enhancement test was 93.62% (95% CI: 82.69–97.66%), with 3 out of 9 KPC and NDM-coproducing strains being missed. By delivering reliable results within 2 h, the R-AST provides an efficient, dependable, and broadly applicable approach to rapid antimicrobial susceptibility testing in clinical settings, potentially mitigating antibiotic misuse and the spread of drug-resistant bacteria.

Graphical Abstract