{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T01:19:23Z","timestamp":1792113563064,"version":"4.4.0"},"reference-count":47,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2024,5,7]],"date-time":"2024-05-07T00:00:00Z","timestamp":1715040000000},"content-version":"vor","delay-in-days":6,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001411","name":"Indian Council of Medical Research","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001411","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Mycoses"],"published-print":{"date-parts":[[2024,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Due to a delay in diagnosis by conventional techniques and high mortality, the development of a standardised and rapid non\u2010culture\u2010based technique is an unmet need in pulmonary, gastrointestinal, and disseminated forms of mucormycosis. Though limited studies have been conducted for molecular diagnosis, there are no established serologic tests for this highly fatal infection.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objective<\/jats:title><jats:p>To develop and evaluate an indirect in\u2010house enzyme\u2010linked immunosorbent assay (ELISA) utilising antigens of <jats:italic>Rhizopus arrhizus<\/jats:italic> for detecting anti\u2010<jats:italic>Rhizopus<\/jats:italic> antibodies (IgG and IgM) in sera of patients with mucormycosis.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We extracted both secretory and mycelial <jats:italic>Rhizopus<\/jats:italic> antigens using standardised protocols. Bradford assay was used for protein quantification. We then standardised an indirect ELISA using <jats:italic>R.\u2009arrhizus<\/jats:italic> mycelial and secretory antigens (10.0\u2009\u03bcg\/mL in bicarbonate buffer pH\u20099.2) for detecting anti\u2010<jats:italic>Rhizopus<\/jats:italic> IgG and IgM antibodies in patient sera. We included patients with mucormycosis, other fungal infections, and healthy controls. Antibody index value (<jats:italic>E<\/jats:italic>\u2010value) was calculated for each patient sample.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Asparagine broth culture filtrate utilising 85% ammonium sulphate salt fractionation and mycelial homogenate grown in yeast extract peptone dextrose (YPD) broth precipitated with trichloroacetic acid (TCA) yielded a large amount of good\u2010quality protein for the assay. We included 55 patients with mucormycosis (rhino\u2010orbito\u2010cerebral mucormycosis [ROCM, <jats:italic>n<\/jats:italic>\u2009=\u200939], pulmonary [<jats:italic>n<\/jats:italic>\u2009=\u200915], gastrointestinal [<jats:italic>n<\/jats:italic>\u2009=\u20091]), 24 with other fungal infections (probable aspergillosis [<jats:italic>n<\/jats:italic>\u2009=\u200914], candidiasis [<jats:italic>n<\/jats:italic>\u2009=\u200910]), and healthy controls (<jats:italic>n<\/jats:italic>\u2009=\u200916). The sensitivity of the antibody test for diagnosing mucormycosis ranged from 83.6\u201392.7% for IgG and 72.7\u201387.3% for IgM, with a specificity of 91.7\u201392.5% for IgG and 80\u201382.5% for IgM. The sera from patients with other fungal infections and healthy individuals did not show significant cross\u2010reactivity.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>The detection of anti\u2010<jats:italic>Rhizopus<\/jats:italic> IgG antibody performed significantly better in comparison to IgM\u2010based ELISA for diagnosing both ROCM (sensitivity of 84.6% vs. 69.2%) and pulmonary cases (86.6% vs. 80.0%). More extensive studies are required to confirm our findings.<\/jats:p><\/jats:sec>","DOI":"10.1111\/myc.13730","type":"journal-article","created":{"date-parts":[[2024,5,7]],"date-time":"2024-05-07T11:48:38Z","timestamp":1715082518000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["A novel indirect <scp>ELISA<\/scp> for serodiagnosis of mucormycosis using antigens from <i>Rhizopus arrhizus<\/i>"],"prefix":"10.1111","volume":"67","author":[{"given":"Hansraj","family":"Choudhary","sequence":"first","affiliation":[{"name":"Indian Council of Medical Research (ICMR)  New Delhi India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4871-8113","authenticated-orcid":false,"given":"Harsimran","family":"Kaur","sequence":"additional","affiliation":[{"name":"Department of Medical Microbiology Postgraduate Institute of Medical Education &amp; Research (PGIMER)  Chandigarh India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shreya","family":"Singh","sequence":"additional","affiliation":[{"name":"Department of Medical Microbiology Dr. B. 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