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<p>Based on the differences in drug resistance characteristics between <i>Candida glabrata</i> and <i>Candida krusei</i> and the clinical need for rapid discrimination, this study established a duplex recombinase polymerase amplification-lateral flow strip (RPA-LFS) detection system using dual-labeled probes. By optimizing ITS2-targeted primers and probes (5′-end labeled with FITC/DIG, 3′-end labeled with biotin) and integrating dual test lines on the strip (streptavidin-T line for directional capture), simultaneous visual detection of both targets was achieved. Performance validation demonstrated: a detection limit of 10 copies/mL and 100 copies/mL, matching qPCR sensitivity; 100% detection rate for 30 target strains (including 12 reference and clinical isolates) with strict discrimination from 8 closely related pathogens (0% cross-reactivity); high concordance with qPCR in 328 clinical samples (sensitivity, specificity, and total concordance all 100%). The system delivers “sample-to-result” output within 30&#xa0;min without complex instrumentation, providing technical support for precise point-of-care discrimination of drug-resistant Candida infections and rational antifungal drug use, particularly in primary healthcare settings and outbreak scenarios.</p>

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Establishment and application of a duplex RPA-LFS detection system for Candida glabrata and Candida krusei

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年2月7日

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  • Lei Wang,
  • Yao Lu,
  • Ting Zhang,
  • Yuanyuan Li,
  • Kun Wang

摘要

Based on the differences in drug resistance characteristics between Candida glabrata and Candida krusei and the clinical need for rapid discrimination, this study established a duplex recombinase polymerase amplification-lateral flow strip (RPA-LFS) detection system using dual-labeled probes. By optimizing ITS2-targeted primers and probes (5′-end labeled with FITC/DIG, 3′-end labeled with biotin) and integrating dual test lines on the strip (streptavidin-T line for directional capture), simultaneous visual detection of both targets was achieved. Performance validation demonstrated: a detection limit of 10 copies/mL and 100 copies/mL, matching qPCR sensitivity; 100% detection rate for 30 target strains (including 12 reference and clinical isolates) with strict discrimination from 8 closely related pathogens (0% cross-reactivity); high concordance with qPCR in 328 clinical samples (sensitivity, specificity, and total concordance all 100%). The system delivers “sample-to-result” output within 30 min without complex instrumentation, providing technical support for precise point-of-care discrimination of drug-resistant Candida infections and rational antifungal drug use, particularly in primary healthcare settings and outbreak scenarios.