{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T04:59:44Z","timestamp":1785387584909,"version":"3.55.0"},"reference-count":50,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T00:00:00Z","timestamp":1785369600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      <jats:italic>Candida tropicalis<\/jats:italic>\n                      (\n                      <jats:italic>C. tropicalis<\/jats:italic>\n                      ), classified as a World Health Organization \u201ccritical priority\u201d pathogen, causes life-threatening invasive infections with high mortality due to diagnostic delays. Therefore, the development of rapid fungal detection platforms is an urgent scientific and clinical priority.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      We designed multiple cross-displacement amplification (MCDA) primers to target the internal transcribed spacer 2 (ITS II) gene of\n                      <jats:italic>C. tropicalis<\/jats:italic>\n                      for specific amplification. Subsequently, the clustered regularly interspaced short palindromic repeats (CRISPR)\/Cas12a-crRNA complex bound to the amplified products, activating its trans-cleavage activity to generate a detectable fluorescent signal. Finally, clinical samples were used to validate the detection results, which were further compared with those obtained using fungal culture and multiplex polymerase chain reaction (Multiplex PCR).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Under optimized conditions, the\n                      <jats:italic>C. tropicalis<\/jats:italic>\n                      -MCDA\u2212CRISPR\/Cas12a assay was completed within approximately 53\u00a0min, with a limit of detection of 30 fg of genomic DNA per reaction. The assay showed no cross\u2212reactivity with non\u2212\n                      <jats:italic>C. tropicalis<\/jats:italic>\n                      pathogens. Clinical validation using 128 specimens demonstrated a sensitivity of 100.0% (95% CI: 93.4\u2013100.0%) and a specificity of 97.3% (95% CI: 90.6\u201399.7%) against a composite reference standard, with near\u2212perfect agreement (\u03ba= 0.968).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>\n                      The\n                      <jats:italic>C. tropicalis<\/jats:italic>\n                      -MCDA-CRISPR\/Cas12a assay is an efficient, accurate, and practical diagnostic tool suitable for use in resource-limited laboratories.\n                    <\/jats:p>\n                  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