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<p>The widespread empirical use of antibiotics significantly worsens bacterial resistance. It is crucial to develop rapid methods for bacterial antibiotic susceptibility testing (AST) that can provide results within a single visit, as this is essential for assessing treatment and improving patient outcomes. However, the gold standard phenotype-based AST methods are limited by complexity and the inability to provide timely AST results within a single visit. Leveraging the hydrogel network’s nano-confined effect on long-chain nucleic acid products for localized amplification, combined with the rapid amplification capability of loop-mediated isothermal amplification (LAMP) technology, we achieved rapid and digital quantification of nucleic acids without relying on microfabrication to microwells or droplets and derived results through artificial intelligence algorithms. This method enables consistent operational steps and timeframes (25-minute antibiotic exposure followed by 16-minute digital nucleic acid quantification) to obtain results for both single and mixed <i>E. coli</i> infections. The entire process takes ~ 45&#xa0;min when used with specimens from urinary tract bacterial infection. H-LAMP (LAMP in hydrogel)based AST demonstrates high consistency with the clinical gold standard of susceptibility testing methods (AUC = 0.98) in 24 patients, using miniaturized and portable devices. This has the potential to advance the clinical management of infectious diseases across a broader spectrum of microorganisms, antibiotics, and sample types, and to improve antimicrobial stewardship.</p>

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Antibiotic Susceptibility Testing from Unprocessed Urine at the Point of Visit

  • 期刊论文

  • 发表日期：

    2026年2月26日

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  • Jidong Wang,
  • Luhua Xin,
  • Zhixin Li,
  • Xi Lu,
  • Jie Lian,
  • Jinxin Zheng,
  • Wenwen Chen

摘要

The widespread empirical use of antibiotics significantly worsens bacterial resistance. It is crucial to develop rapid methods for bacterial antibiotic susceptibility testing (AST) that can provide results within a single visit, as this is essential for assessing treatment and improving patient outcomes. However, the gold standard phenotype-based AST methods are limited by complexity and the inability to provide timely AST results within a single visit. Leveraging the hydrogel network’s nano-confined effect on long-chain nucleic acid products for localized amplification, combined with the rapid amplification capability of loop-mediated isothermal amplification (LAMP) technology, we achieved rapid and digital quantification of nucleic acids without relying on microfabrication to microwells or droplets and derived results through artificial intelligence algorithms. This method enables consistent operational steps and timeframes (25-minute antibiotic exposure followed by 16-minute digital nucleic acid quantification) to obtain results for both single and mixed E. coli infections. The entire process takes ~ 45 min when used with specimens from urinary tract bacterial infection. H-LAMP (LAMP in hydrogel)based AST demonstrates high consistency with the clinical gold standard of susceptibility testing methods (AUC = 0.98) in 24 patients, using miniaturized and portable devices. This has the potential to advance the clinical management of infectious diseases across a broader spectrum of microorganisms, antibiotics, and sample types, and to improve antimicrobial stewardship.