{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T23:57:10Z","timestamp":1787356630426,"version":"build-2736575974"},"reference-count":37,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2021,10,23]],"date-time":"2021-10-23T00:00:00Z","timestamp":1634947200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81902043"],"award-info":[{"award-number":["81902043"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100018537","name":"National Science and Technology Major Project","doi-asserted-by":"publisher","award":["2018ZX10712001"],"award-info":[{"award-number":["2018ZX10712001"]}],"id":[{"id":"10.13039\/501100018537","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100018537","name":"National Science and Technology Major Project","doi-asserted-by":"publisher","award":["2020YFC2005401"],"award-info":[{"award-number":["2020YFC2005401"]}],"id":[{"id":"10.13039\/501100018537","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Mycoses"],"published-print":{"date-parts":[[2021,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Objective<\/jats:title><jats:p>To develop a comprehensive diagnostic system for mucormycosis from formalin\u2010fixed paraffin\u2010embedded tissues, consisting of own\u2010designed real\u2010time polymerase chain reaction (PCR) assays, fluorescence in situ hybridisation, and immunohistochemical staining.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We designed 11 primers and probes for specific real\u2010time PCR assays based on genome sequences, and validated the specificity by <jats:italic>Aspergillus, Fusarium, Scedosporium, Lomentospora, Cryptococcus<\/jats:italic> and <jats:italic>Candida<\/jats:italic> species. Formalin\u2010fixed paraffin\u2010embedded (FFPE) tissues from forty\u2010four mouse model infected by above fungi were collected and extracted DNA by laser capture microdissection (LCM) and direct extraction methods for real\u2010time PCR assays. In addition, seventeen clinical specimens histopathologically proven for mucormycosis were included for specific detection with the new diagnostic system.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The real\u2010time PCR assays allowed detection of a minimum of 10\u00a0CFU\/ml equivalent gDNA of each species. No cross\u2010reaction with gDNA among species was noted. From mouse model specimens, the sensitivity of real\u2010time PCR in samples extracted with LCM versus direct extraction method was 100% versus 91.43% at Mucorales level and 80% versus 45.71% at species level, respectively. The specificity was 100%. From clinical samples, LCM combined with real\u2010time PCR can test 88.24% (15\/17) of Mucorales. Sensitivities of fluorescence in situ hybridisation (FISH) and immunohistochemical staining (IHC) were 70.59% and 41.18%, respectively. Combined LCM\u2010RT\u2010PCR, FISH and IHC yielded positive results in all samples.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>The combination diagnostic system we developed is a culture\u2010independent and robust method which enables rapid species identification from FFPE tissues for timely diagnosis of mucormycosis.<\/jats:p><\/jats:sec>","DOI":"10.1111\/myc.13382","type":"journal-article","created":{"date-parts":[[2021,10,21]],"date-time":"2021-10-21T16:30:40Z","timestamp":1634833840000},"page":"1460-1470","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["The use of combined PCR, fluorescence in situ hybridisation and immunohistochemical staining to diagnose mucormycosis from formalin\u2010fixed paraffin\u2010embedded tissues"],"prefix":"10.1111","volume":"64","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4040-0530","authenticated-orcid":false,"given":"Xiao","family":"Liu","sequence":"first","affiliation":[{"name":"Department of Dermatology and Venerology Peking University First Hospital Beijing China"},{"name":"Research Center for Medical Mycology Peking University Beijing China"},{"name":"National Clinical Research Center for Skin and Immune Diseases Beijing China"},{"name":"Beijing Key Laboratory of Molecular Diagnosis of Dermatoses Peking University First Hospital Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1644-6942","authenticated-orcid":false,"given":"Yinggai","family":"Song","sequence":"additional","affiliation":[{"name":"Department of Dermatology and Venerology Peking University First Hospital Beijing China"},{"name":"Research Center for Medical Mycology Peking University Beijing China"},{"name":"National Clinical Research Center for Skin and Immune Diseases Beijing China"},{"name":"Beijing Key Laboratory of Molecular Diagnosis of Dermatoses Peking University First Hospital Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruoyu","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Dermatology and Venerology Peking University First Hospital Beijing China"},{"name":"Research Center for Medical Mycology Peking University Beijing China"},{"name":"National Clinical Research Center for Skin and Immune Diseases Beijing China"},{"name":"Beijing Key Laboratory of Molecular Diagnosis of Dermatoses Peking University First Hospital Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2021,10,23]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/coa.13175"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1093\/mmy\/myx017"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1469-0691.2009.02978.x"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1111\/myc.13353"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(19)30312-3"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmi.2018.07.011"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11046-019-00380-5"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.00746-18"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1309\/7KU5XWURYM0151YN"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/ciz1008"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.06133-11"},{"issue":"4","key":"e_1_2_9_13_1","first-page":"385","article-title":"Detection of mucormycetes and other pathogenic fungi in formalin fixed paraffin embedded and fresh tissues using the extended region of 28S rDNA","volume":"55","author":"Gade L","year":"2017","journal-title":"Med Mycol"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.00256-11"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1093\/mmy\/myy041"},{"issue":"11","key":"e_1_2_9_16_1","first-page":"1255","article-title":"Laser capture microdissection: big data from small samples","volume":"30","author":"Datta S","year":"2015","journal-title":"Histol Histopathol"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1080\/22221751.2021.1889931"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1099\/jmm.0.000375"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.00289-18"},{"issue":"4","key":"e_1_2_9_20_1","first-page":"ofw190","article-title":"Is it time to include CT \"reverse halo sign\" and qPCR targeting Mucorales in serum to EORTC\u2010MSG criteria for the diagnosis of pulmonary mucormycosis in leukemia patients? 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