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<p>Urinary tract infections (UTIs) require rapid pathogen identification at the point of care (POCT) to enable timely therapy and reduce empirical antibiotic use. Here, we report an autosampling microfluidic platform that integrates a vacuum-driven PDMS chip with FEN1-aided recombinase polymerase amplification (FARPA), a syringe-filter enrichment module, and a rapid lysis protocol (1-min ultrasonication followed by 2-min thermal lysis at 98˚Ϲ). The round chip partitions each sample into five reaction chambers with preloaded primers/probes, enabling parallel detection of five common UTI pathogens (<i>Escherichia coli</i>,<i> Klebsiella pneumoniae</i>,<i> Acinetobacter baumannii</i>,<i> Enterococcus faecium</i>,<i> and Enterococcus faecalis</i>) across five samples in a single run. Using DNA templates, the chip assay detects down to 5 copies per reaction. With syringe-filter enrichment and rapid lysis, the system achieves the sample-level limit of detection of 10^2 CFU/mL (in urine matrix) and completes the sample-to-answer workflow within 55&#xa0;min using a battery-powered handheld fluorescence reader. In a clinical study of 299 specimens (urine, ascitic fluid, pleural effusion, abdominal drainage, and bladder-puncture urine), the chip-FARPA results showed 91.1% sensitivity (72/79 culture-positive detected) and 100% specificity versus bacterial culture, supporting a portable and low-cost POCT solution for on-site UTI pathogen screening.&gt;</p> Graphical abstract <p></p>

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An autosampling microfluidic chip enables FEN1-aided recombinase polymerase amplification to rapidly detect multiple urinary tract infection pathogens on-site

  • 期刊论文

  • 发表日期：

    2026年4月7日

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  • Long Chen,
  • Liying Feng,
  • Ying Wang,
  • Jieyu Zhang,
  • Chen Chen,
  • XuePing Ma,
  • Haiping Wu,
  • Hao Tang,
  • HuiJie Yue,
  • Guohua Zhou

摘要

Urinary tract infections (UTIs) require rapid pathogen identification at the point of care (POCT) to enable timely therapy and reduce empirical antibiotic use. Here, we report an autosampling microfluidic platform that integrates a vacuum-driven PDMS chip with FEN1-aided recombinase polymerase amplification (FARPA), a syringe-filter enrichment module, and a rapid lysis protocol (1-min ultrasonication followed by 2-min thermal lysis at 98˚Ϲ). The round chip partitions each sample into five reaction chambers with preloaded primers/probes, enabling parallel detection of five common UTI pathogens (Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, Enterococcus faecium, and Enterococcus faecalis) across five samples in a single run. Using DNA templates, the chip assay detects down to 5 copies per reaction. With syringe-filter enrichment and rapid lysis, the system achieves the sample-level limit of detection of 10^2 CFU/mL (in urine matrix) and completes the sample-to-answer workflow within 55 min using a battery-powered handheld fluorescence reader. In a clinical study of 299 specimens (urine, ascitic fluid, pleural effusion, abdominal drainage, and bladder-puncture urine), the chip-FARPA results showed 91.1% sensitivity (72/79 culture-positive detected) and 100% specificity versus bacterial culture, supporting a portable and low-cost POCT solution for on-site UTI pathogen screening.>

Graphical abstract