{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T13:39:49Z","timestamp":1792676389532,"version":"4.5.0"},"reference-count":23,"publisher":"American Society for Microbiology","license":[{"start":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T00:00:00Z","timestamp":1792627200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T00:00:00Z","timestamp":1792627200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R01AI150823"],"award-info":[{"award-number":["R01AI150823"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"name":"UKRI Future Leaders Fellowship","award":["MR\/V026097\/1"],"award-info":[{"award-number":["MR\/V026097\/1"]}]},{"name":"Sight Research UK Translational Research Award","award":["TRN003"],"award-info":[{"award-number":["TRN003"]}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"abstract":"ABSTRACT<\/jats:title>\n \n \n \n KOH microscopy is a routine method to determine the presence of fungal elements in infected tissue. Although widely used, the procedure requires trained personnel and the infrastructure needed for microscopy. An alternative approach is the extraction of fungal antigens and detection by immunoassay. In this study, we used oxidative release with sodium hypochlorite to extract mannan from fungal cells. Quenching of free sodium hypochlorite was done using an excess of bovine serum albumin or sodium thiosulfate. Quenching reagents were incorporated into sample pads of lateral flow immunoassays (LFIAs), followed by detection of released mannan using mannan-specific monoclonal antibodies coupled to colloidal gold on the LFIA platform. Cells of\n Candida<\/jats:italic>\n spp. or\n Aspergillus<\/jats:italic>\n spp. were used to evaluate the extraction\/quenching steps. The results showed that sodium thiosulfate was particularly well suited for hands-free, in-strip neutralization. The analytical sensitivity (minimum limit of detection, LOD) of the\n Candida<\/jats:italic>\n LFIA was 67 ng\/mL of formaldehyde-inactivated, whole\n Candida albicans<\/jats:italic>\n cells. The LOD of the\n Aspergillus<\/jats:italic>\n LFIA was 250 ng\/mL of formaldehyde-inactivated, whole\n Aspergillus fumigatus<\/jats:italic>\n cells. Proof-of-concept for oxidative release of mannan followed by detection of antigen by LFIA was provided using an\n ex vivo<\/jats:italic>\n porcine corneal model of fungal keratitis. The results demonstrated that the test is a rapid two-step, specimen-to-result protocol suitable for point-of-care use. Such a diagnostic would be particularly valuable in settings that lack access to the trained personnel or infrastructure needed for KOH microscopy.\n <\/jats:p>\n \n IMPORTANCE<\/jats:title>\n \n KOH microscopy is widely used to detect fungal infection, but it requires trained personnel and microscopy infrastructure. An alternative approach is extraction of antigens from fungal cells within infected tissue and detection by lateral flow immunoassay. In this study, we developed a <20 min, specimen-to-result protocol suitable for point-of-care use that combines (i) oxidative release via sodium hypochlorite to rapidly extract fungal mannan from specimens, (ii) on-strip (hands-free) quenching of sodium hypochlorite with sodium thiosulfate or bovine serum albumin, and (iii) visual detection of positive signal via novel, mannan-specific monoclonal antibodies coupled to colloidal gold. Analytical performance of the\n Candida<\/jats:italic>\n and\n Aspergillus<\/jats:italic>\n tests was evaluated with cultured fungal cells. Clinical proof-of-concept for each assay was provided using an\n ex vivo<\/jats:italic>\n porcine corneal model of fungal keratitis. This diagnostic strategy would be particularly valuable in settings that lack access to trained personnel or the infrastructure needed for KOH microscopy.\n <\/jats:p>\n <\/jats:sec>\n <\/jats:sec>","DOI":"10.1128\/spectrum.02253-26","type":"journal-article","created":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T13:00:37Z","timestamp":1792674037000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["A microscopy-free approach for detection of fungal elements in tissue using mannan immunoassay"],"prefix":"10.1128","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-9109-3087","authenticated-orcid":true,"given":"Audrey M.","family":"Rank","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01keh0577","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology and Immunology, University of Nevada, Reno School of Medicine","place":["Reno, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2320-2222","authenticated-orcid":true,"given":"Amanda R.","family":"Burnham-Marusich","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01keh0577","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology and Immunology, University of Nevada, Reno School of Medicine","place":["Reno, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kathleen R.","family":"Zayac","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01keh0577","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology and Immunology, University of Nevada, Reno School of Medicine","place":["Reno, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Menen G.","family":"Ashagrie","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01keh0577","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology and Immunology, University of Nevada, Reno School of Medicine","place":["Reno, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lawrence","family":"Mariano","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01keh0577","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology and Immunology, University of Nevada, Reno School of Medicine","place":["Reno, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4397-4019","authenticated-orcid":false,"given":"Sheelagh","family":"Duncan","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/05wcr1b38","id-type":"ROR","asserted-by":"publisher"}],"name":"Centre for Inflammation Research, Institute for Regeneration and Repair, University of Edinburgh","place":["Edinburgh, United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3209-9490","authenticated-orcid":false,"given":"Bethany","family":"Mills","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/05wcr1b38","id-type":"ROR","asserted-by":"publisher"}],"name":"Centre for Inflammation Research, Institute for Regeneration and Repair, University of Edinburgh","place":["Edinburgh, United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1477-3261","authenticated-orcid":false,"given":"Thomas R.","family":"Kozel","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01keh0577","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Microbiology and Immunology, University of Nevada, Reno School of Medicine","place":["Reno, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","published-online":{"date-parts":[[2026,10,22]]},"reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/ciae104"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/mSphere.00094-18"},{"key":"e_1_3_4_4_2","volume-title":"Platelia Aspergillus Ag Ref 62794","author":"Bio-Rad","year":"2013","unstructured":"Bio-Rad. 2013. 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