{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,14]],"date-time":"2025-12-14T08:27:15Z","timestamp":1765700835561},"reference-count":19,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2002,3,1]],"date-time":"2002-03-01T00:00:00Z","timestamp":1014940800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Clin Vaccine Immunol"],"published-print":{"date-parts":[[2002,3]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Two PCR protocols targeting the 18S rRNA gene of\n            <jats:italic>Cryptococcus neoformans<\/jats:italic>\n            were established, compared, and evaluated in murine cryptococcal meningitis. One protocol was designed as a nested PCR to be performed in conventional block thermal cyclers. The other protocol was designed as a quantitative single-round PCR adapted to LightCycler technology. One hundred brain homogenates and dilutions originating from 20 ICR mice treated with different azoles were examined. A fungal burden of 3 \u00d7 10\n            <jats:sup>1<\/jats:sup>\n            to 2.9 \u00d7 10\n            <jats:sup>4<\/jats:sup>\n            CFU per mg of brain tissue was determined by quantitative culture. Specific PCR products were amplified by the conventional and the LightCycler methods in 86 and 87 samples, respectively, with products identified by DNA sequencing and real-time fluorescence detection. An analytical sensitivity of 1 CFU of\n            <jats:italic>C. neoformans<\/jats:italic>\n            per mg of brain tissue and less than 10 CFU per volume used for extraction was observed for both PCR protocols, while homogenates of 70 organs from mice infected with other fungi were PCR negative. Specificity testing was performed with genomic DNA from 31 hymenomycetous fungal species and from the ustilaginomycetous yeast\n            <jats:italic>Malassezia furfur<\/jats:italic>\n            , which are phylogenetically related to\n            <jats:italic>C. neoformans<\/jats:italic>\n            . Twenty-four strains, including species of human skin flora like\n            <jats:italic>M. furfur<\/jats:italic>\n            and\n            <jats:italic>Trichosporon<\/jats:italic>\n            spp., were PCR negative. Amplification was observed with\n            <jats:italic>Cryptococcus amylolentus<\/jats:italic>\n            ,\n            <jats:italic>Filobasidiella depauperata<\/jats:italic>\n            ,\n            <jats:italic>Cryptococcus laurentii<\/jats:italic>\n            , and five species unrelated to clinical specimens. LightCycler PCR products from\n            <jats:italic>F. depauperata<\/jats:italic>\n            and\n            <jats:italic>Trichosporon faecale<\/jats:italic>\n            could be clearly discriminated by melting curve analysis. The sensitive and specific nested PCR assay as well as the rapid and quantitative LightCycler PCR assay might be useful for the diagnosis and monitoring of human cryptococcal infections.\n          <\/jats:p>","DOI":"10.1128\/cdli.9.2.461-469.2002","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:58:35Z","timestamp":1027763915000},"page":"461-469","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":7,"title":["Detection of\n            <i>Cryptococcus neoformans<\/i>\n            DNA in Tissue Samples by Nested and Real-Time PCR Assays"],"prefix":"10.1128","volume":"9","author":[{"given":"Ralf","family":"Bialek","sequence":"first","affiliation":[{"name":"Institute for Tropical Medicine, University Hospital Tu\u0308bingen"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Weiss","sequence":"additional","affiliation":[{"name":"Botanical Institute, University of Tu\u0308bingen, Tu\u0308bingen"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kubrom","family":"Bekure-Nemariam","sequence":"additional","affiliation":[{"name":"Institute for Tropical Medicine, University Hospital Tu\u0308bingen"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Laura K.","family":"Najvar","sequence":"additional","affiliation":[{"name":"University of Texas Health Science Center at San Antonio, San Antonio, Texas"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria B.","family":"Alberdi","sequence":"additional","affiliation":[{"name":"Institute of Medical Microbiology, University of Regensburg, Regensburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John R.","family":"Graybill","sequence":"additional","affiliation":[{"name":"University of Texas Health Science Center at San Antonio, San Antonio, Texas"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Udo","family":"Reischl","sequence":"additional","affiliation":[{"name":"Institute of Medical Microbiology, University of Regensburg, Regensburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.38.8.2940-2942.2000"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.39.4.1506-1509.2001"},{"key":"e_1_3_2_4_2","doi-asserted-by":"crossref","first-page":"1674","DOI":"10.1128\/JCM.33.6.1674-1675.1995","volume":"33","year":"1995","unstructured":"Blevins, L. 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