{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,16]],"date-time":"2026-04-16T00:56:52Z","timestamp":1776301012123,"version":"3.50.1"},"reference-count":24,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2022,8,11]],"date-time":"2022-08-11T00:00:00Z","timestamp":1660176000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"MGM foundation, Bhubaneswar, Odisha, India","award":["25050"],"award-info":[{"award-number":["25050"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JoF"],"abstract":"<jats:p>Early diagnosis and treatment of rhino-orbital-cerebral mucormycosis (ROCM) are crucial. Potassium hydroxide with Calcofluorwhite (KOH + CFW) smears can demonstrate the fungal hyphae, but mixed infections caused by both mucorales and non-mucorales pose a diagnostic challenge. Polymerase chain reaction (PCR) can detect mixed infections and differentiate mucorales from non-mucorales. This study aimed to evaluate the utility of a single reaction PCR in the diagnosis of ROCM and the efficacy of nasal biopsy and endonasal swab in the detection of fungus. Sixty-six clinical samples were collected from 33 patients and were subjected to KOH + CFW smear, culture and PCR. PCR was performed using pan-fungal primers targeting the 28S large subunit rRNA gene, and the amplified products were further sequenced to identify the fungi. KOH + CFW smear, culture and PCR detected mucorales in 54.6%, 27.3% and 63.6% patients, respectively. PCR detected mixed infection in 51.5% patients compared to 9.1% by KOH + CFW smear. PCR detected fungus in 90% of nasal biopsies and 77.8% of endonasal swabs. Rhizopus spp. was the most common fungi identified in 43.2% of PCR-positive samples. PCR is effective in detecting mixed infection and in the diagnosis of ROCM. Nasal biopsies had better fungal detection rates than endonasal swabs.<\/jats:p>","DOI":"10.3390\/jof8080844","type":"journal-article","created":{"date-parts":[[2022,8,11]],"date-time":"2022-08-11T21:15:05Z","timestamp":1660252505000},"page":"844","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Diagnostic Performance and Clinical Utility of Conventional PCR Assay in Early Diagnosis of COVID-19 Associated Rhino-Orbito-Cerebral Mucormycosis"],"prefix":"10.3390","volume":"8","author":[{"given":"Samir","family":"Mohapatra","sequence":"first","affiliation":[{"name":"Department of Ophthalmic Plastic and Reconstructive Surgery Service, L V Prasad Eye Institute, Mithu Tulsi Chanrai Campus, Bhubaneswar 751024, Odisha, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manas Ranjan","family":"Barik","sequence":"additional","affiliation":[{"name":"Ocular Microbiology Service, L V Prasad Eye Institute, Mithu Tulsi Chanrai Campus, Bhubaneswar 751024, Odisha, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Suryasnata","family":"Rath","sequence":"additional","affiliation":[{"name":"Department of Ophthalmic Plastic and Reconstructive Surgery Service, L V Prasad Eye Institute, Mithu Tulsi Chanrai Campus, Bhubaneswar 751024, Odisha, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9979-264X","authenticated-orcid":false,"given":"Savitri","family":"Sharma","sequence":"additional","affiliation":[{"name":"Jhaveri Microbiology Centre, L V Prasad Eye Institute, Kallam Anji Reddy Campus, Hyderabad 500034, Telangana, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Archisman","family":"Mohapatra","sequence":"additional","affiliation":[{"name":"Department of Biostatistics, Generating Research Insights for Development (GRID) Council, Noida 201307, Delhi NCR, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1861-6113","authenticated-orcid":false,"given":"Sharmistha","family":"Behera","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, VSSIMSAR, Burla 768017, Odisha, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3580-0211","authenticated-orcid":false,"given":"Souvagini","family":"Acharya","sequence":"additional","affiliation":[{"name":"Department of Ear, Nose and Throat, VSSIMSAR, Burla 768017, Odisha, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dipti Ranjan","family":"Pattjoshi","sequence":"additional","affiliation":[{"name":"Department of Ear, Nose and Throat, SCB Medical College and Hospital, Cuttack 753007, Odisha, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rajesh Kumar","family":"Padhi","sequence":"additional","affiliation":[{"name":"Department of Ear, Nose and Throat, Sparsh Hospitals Pvt Ltd., Bhubaneswar 751007, Odisha, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4638-2836","authenticated-orcid":false,"given":"Himansu Sekhar","family":"Behera","sequence":"additional","affiliation":[{"name":"Ocular Microbiology Service, L V Prasad Eye Institute, Mithu Tulsi Chanrai Campus, Bhubaneswar 751024, Odisha, India"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,8,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1002","DOI":"10.4103\/ijo.IJO_3763_20","article-title":"COVID-19 and orbital mucormycosis","volume":"69","author":"Sarkar","year":"2021","journal-title":"Indian J. Ophthalmol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1361","DOI":"10.4103\/ijo.IJO_1165_21","article-title":"Code Mucor: Guidelines for the Diagnosis, Staging and Management of Rhino-Orbito-Cerebral Mucormycosis in the Setting of COVID-19","volume":"69","author":"Honavar","year":"2021","journal-title":"Indian J. Ophthalmol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"507064","DOI":"10.3389\/fendo.2020.507064","article-title":"Prevalence of Diabetes and Its Determinants in the Young Adults Indian Population-Call for Yoga Intervention","volume":"11","author":"Nagarathna","year":"2020","journal-title":"Front. Endocrinol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"100722","DOI":"10.1016\/j.jemep.2021.100722","article-title":"Delta variant and black fungal invasion: A bidirectional assault might worsen the massive second\/third stream of COVID-19 outbreak in South-Asia","volume":"19","author":"Bari","year":"2021","journal-title":"Ethic-Med. Public Health"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"e93","DOI":"10.1016\/S2213-2600(21)00312-X","article-title":"Implications of the second wave of COVID-19 in India","volume":"9","author":"Asrani","year":"2021","journal-title":"Lancet Respir. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1007\/s10096-006-0107-1","article-title":"Zygomycosis: The re-emerging fungal infection","volume":"25","author":"Chayakulkeeree","year":"2006","journal-title":"Eur. J. Clin. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"47","DOI":"10.4103\/jacm.jacm_2_19","article-title":"Mixed opportunistic infection with Mucor, Aspergillus and Candida in oculo-rhino-cerebral mycosis: An uncommon case","volume":"21","author":"Pandey","year":"2019","journal-title":"J. Acad. Clin. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1093\/mmy\/myy060","article-title":"A prospective multicenter study on mucormycosis in India: Epidemiology, diagnosis, and treatment","volume":"57","author":"Prakash","year":"2019","journal-title":"Med. Mycol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Sipsas, N.V., Gamaletsou, M.N., Anastasopoulou, A., and Kontoyiannis, D.P. (2018). Therapy of Mucormycosis. J. Fungi, 4.","DOI":"10.3390\/jof4030090"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1124","DOI":"10.1099\/jmm.0.000560","article-title":"Molecular diagnosis of rhino-orbito-cerebral mucormycosis from fresh tissue samples","volume":"66","author":"Zaman","year":"2017","journal-title":"J. Med. Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"630","DOI":"10.1111\/ceo.13939","article-title":"Simple polymerase chain reaction assay to differentiate between fungal and Pythium insidiosum keratitis","volume":"49","author":"Behera","year":"2021","journal-title":"Clin. Exp. Ophthalmol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1080\/08820538.2021.1946094","article-title":"The ROC Staging System for COVID-related Rhino-Orbital-Cerebral Mucormycosis","volume":"37","author":"Naik","year":"2021","journal-title":"Semin. Ophthalmol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2913","DOI":"10.1128\/jcm.33.11.2913-2919.1995","article-title":"Molecular probes for diagnosis of fungal infections","volume":"33","author":"Sandhu","year":"1995","journal-title":"J. Clin. Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e405","DOI":"10.1016\/S1473-3099(19)30312-3","article-title":"Mucormycosis ECMM MSG Global Guideline Writing Group. Global guideline for the diagnosis and management of mucormycosis: An initiative of the European Confederation of Medical Mycology in cooperation with the Mycoses Study Group Education and Research Consortium","volume":"19","author":"Cornely","year":"2019","journal-title":"Lancet Infect. Dis."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Aranjani, J.M., Manuel, A., Razack, H.I.A., and Mathew, S.T. (2021). COVID-19\u2013associated mucormycosis: Evidence-based critical review of an emerging infection burden during the pandemic\u2019s second wave in India. PLoS Negl. Trop. Dis., 15.","DOI":"10.1371\/journal.pntd.0009921"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e1005491","DOI":"10.1371\/journal.ppat.1005491","article-title":"Filamentous Fungi in Respiratory Infections. What Lies Beyond Aspergillosis and Mucormycosis?","volume":"12","author":"Chowdhary","year":"2016","journal-title":"PLOS Pathog."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1670","DOI":"10.4103\/ijo.IJO_1565_21","article-title":"Epidemiology, clinical profile, management, and outcome of COVID-19-associated rhino-orbital-cerebral mucormycosis in 2826 patients in India\u2014Collaborative OPAI-IJO Study on Mucormycosis in COVID-19 (COSMIC), Report 1","volume":"69","author":"Sen","year":"2021","journal-title":"Indian J. Ophthalmol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"JCM0123021","DOI":"10.1128\/JCM.01230-21","article-title":"Epidemiology and Antifungal Susceptibilities of Mucoralean Fungi in Clinical Samples from the United States","volume":"59","author":"Badali","year":"2021","journal-title":"J. Clin. Microbiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1080\/13693780500464930","article-title":"The rising trend of invasive zygomycosis in patients with uncontrolled diabetes mellitus","volume":"44","author":"Chakrabarti","year":"2006","journal-title":"Med. Mycol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1970","DOI":"10.4103\/ijo.IJO_1143_21","article-title":"COVID-19, steroids, and mucormycosis: What an ophthalmologist should know","volume":"69","author":"Tandon","year":"2021","journal-title":"Indian J. Ophthalmol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"S35","DOI":"10.1093\/cid\/cir880","article-title":"A global analysis of mucormycosis in France: The RetroZygo Study (2005\u20132007)","volume":"54","author":"Lanternier","year":"2012","journal-title":"Clin. Infect. Dis."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"573","DOI":"10.1136\/pgmj.2008.076463","article-title":"Invasive zygomycosis in India: Experience in a tertiary care hospital","volume":"85","author":"Chakrabarti","year":"2009","journal-title":"Postgrad. Med J."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"S93","DOI":"10.1093\/mmy\/myx101","article-title":"Challenges in the diagnosis and treatment of mucormycosis","volume":"56","author":"Skiada","year":"2018","journal-title":"Med Mycol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"863","DOI":"10.1128\/JCM.02965-12","article-title":"Utility of PCR in Diagnosis of Invasive Fungal Infections: Real-Life Data from a Multicenter Study","volume":"51","author":"Mutschlechner","year":"2013","journal-title":"J. Clin. Microbiol."}],"container-title":["Journal of Fungi"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2309-608X\/8\/8\/844\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:07:29Z","timestamp":1760141249000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2309-608X\/8\/8\/844"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,8,11]]},"references-count":24,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2022,8]]}},"alternative-id":["jof8080844"],"URL":"https:\/\/doi.org\/10.3390\/jof8080844","relation":{},"ISSN":["2309-608X"],"issn-type":[{"value":"2309-608X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,8,11]]}}}