Scholarlink

<p><i>Enterococcus faecium</i> and <i>Enterococcus faecalis</i>, ranked among the top three global nosocomial pathogens (WHO data), pose escalating threats due to multidrug resistance (particularly vancomycin-resistant strains, VRE) and foodborne transmission. Conventional detection relies on time-consuming culture-based typing (ISO 7899-2; 48–72&#xa0;h) with &gt; 12% false-negative rates, while molecular methods like qPCR require sophisticated instrumentation, limiting field deployment. This study developed a rapid dual recombinase polymerase amplification-lateral flow strip (RPA-LFS) assay for simultaneous detection of <i>E. faecium</i> and <i>E. faecalis</i>, addressing critical bottlenecks of prolonged turnaround time, species differentiation challenges, and equipment dependency. Species-specific primers and probes were designed targeting conserved genomic regions. Probes featured 5’-FITC/DIG reporter groups and amplification products incorporated 3’-biotin modifications, enabling THF cleavage-mediated signal release. Following optimization (37&#xa0;°C, 20&#xa0;min), results were visually interpreted via LFS within 10&#xa0;min. Key findings include: Sensitivity: 10 CFU/mL (<i>E. faecalis</i>) and 10<sup>2</sup> CFU/mL (<i>E. faecium</i>), maintained in complex matrices; Specificity: 100% (no cross-reactivity with 30 non-target strains); Clinical validation: 100% sensitivity and specificity (<i>n</i> = 435 sputum isolates), significantly outperforming culture methods. This assay achieves equipment-free species differentiation within 40&#xa0;min, providing a transformative tool for precision antimicrobial therapy, food safety monitoring, and frontline antimicrobial resistance (AMR) containment.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Development and application of a rapid dual RPA-LFS assay for simultaneous detection of Enterococcus faecalis and Enterococcus faecium

  • 期刊论文

  • 发表日期：

    2025年11月28日

查看全文
  • Yang Wang,
  • Zhongfang Dai,
  • Wenjun Zhu,
  • Lei Wang,
  • Hongchun Li

摘要

Enterococcus faecium and Enterococcus faecalis, ranked among the top three global nosocomial pathogens (WHO data), pose escalating threats due to multidrug resistance (particularly vancomycin-resistant strains, VRE) and foodborne transmission. Conventional detection relies on time-consuming culture-based typing (ISO 7899-2; 48–72 h) with > 12% false-negative rates, while molecular methods like qPCR require sophisticated instrumentation, limiting field deployment. This study developed a rapid dual recombinase polymerase amplification-lateral flow strip (RPA-LFS) assay for simultaneous detection of E. faecium and E. faecalis, addressing critical bottlenecks of prolonged turnaround time, species differentiation challenges, and equipment dependency. Species-specific primers and probes were designed targeting conserved genomic regions. Probes featured 5’-FITC/DIG reporter groups and amplification products incorporated 3’-biotin modifications, enabling THF cleavage-mediated signal release. Following optimization (37 °C, 20 min), results were visually interpreted via LFS within 10 min. Key findings include: Sensitivity: 10 CFU/mL (E. faecalis) and 102 CFU/mL (E. faecium), maintained in complex matrices; Specificity: 100% (no cross-reactivity with 30 non-target strains); Clinical validation: 100% sensitivity and specificity (n = 435 sputum isolates), significantly outperforming culture methods. This assay achieves equipment-free species differentiation within 40 min, providing a transformative tool for precision antimicrobial therapy, food safety monitoring, and frontline antimicrobial resistance (AMR) containment.