{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,31]],"date-time":"2025-10-31T07:53:20Z","timestamp":1761897200694},"reference-count":35,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[2000,4,1]],"date-time":"2000-04-01T00:00:00Z","timestamp":954547200000},"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":["J Clin Microbiol"],"published-print":{"date-parts":[[2000,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            A PCR-based assay was developed to detect and identify medically important yeasts in clinical samples. Using a previously described set of primers (G. Morace et al., J. Clin. Microbiol. 35:667\u2013672, 1997), we amplified a fragment of the\n            <jats:italic>ERG11<\/jats:italic>\n            gene for cytochrome P-450 lanosterol 14\u03b1-demethylase, a crucial enzyme in the biosynthesis of ergosterol. The PCR product was analyzed in a reverse cross blot hybridization assay with species-specific probes directed to a target region of the\n            <jats:italic>ERG11<\/jats:italic>\n            gene of\n            <jats:italic>Candida albicans<\/jats:italic>\n            (pCal),\n            <jats:italic>C. guilliermondii<\/jats:italic>\n            (pGui),\n            <jats:italic>C.<\/jats:italic>\n            (\n            <jats:italic>Torulopsis<\/jats:italic>\n            )\n            <jats:italic>glabrata<\/jats:italic>\n            (pGla),\n            <jats:italic>C. kefyr<\/jats:italic>\n            (pKef),\n            <jats:italic>C. krusei<\/jats:italic>\n            (pKru),\n            <jats:italic>C. parapsilosis<\/jats:italic>\n            (pPar),\n            <jats:italic>C. tropicalis<\/jats:italic>\n            (pTro), the newly described species\n            <jats:italic>C. dubliniensis<\/jats:italic>\n            (pDub),\n            <jats:italic>Saccharomyces cerevisiae<\/jats:italic>\n            (pSce), and\n            <jats:italic>Cryptococcus neoformans<\/jats:italic>\n            (pCry). The PCR-reverse cross blot hybridization assay correctly identified multiple isolates of each species tested. No cross-hybridization was detected with any other fungal, bacteria, or human DNAs tested. The method was tested against conventional identification on 140 different clinical samples, including blood and cerebrospinal fluid, from patients with suspected fungal infections. The results agreed with those of culture and phenotyping for all but six specimens (two of which grew yeasts not included in the PCR panel of probes and four in which PCR positivity-culture negativity was justified by clinical findings). Species identification time was reduced from a mean of 4 days with conventional identification to 7 h with the molecular method. The PCR-reverse cross blot hybridization assay is a rapid method for the direct detection and identification of yeasts in clinical samples.\n          <\/jats:p>","DOI":"10.1128\/jcm.38.4.1609-1614.2000","type":"journal-article","created":{"date-parts":[[2019,12,31]],"date-time":"2019-12-31T17:55:23Z","timestamp":1577814923000},"page":"1609-1614","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":30,"title":["Reverse Cross Blot Hybridization Assay for Rapid Detection of PCR-Amplified DNA from\n            <i>Candida<\/i>\n            Species,\n            <i>Cryptococcus neoformans<\/i>\n            , and\n            <i>Saccharomyces cerevisiae<\/i>\n            in Clinical Samples"],"prefix":"10.1128","volume":"38","author":[{"given":"Brunella","family":"Posteraro","sequence":"first","affiliation":[{"name":"<!--label omitted: 1-->Istituto di Microbiologia, Universita\u0300Cattolica del Sacro Cuore, Rome,1 and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maurizio","family":"Sanguinetti","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Istituto di Microbiologia, Universita\u0300Cattolica del Sacro Cuore, Rome,1 and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luca","family":"Masucci","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Istituto di Microbiologia, Universita\u0300Cattolica del Sacro Cuore, Rome,1 and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lucio","family":"Romano","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Istituto di Microbiologia, Universita\u0300Cattolica del Sacro Cuore, Rome,1 and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giulia","family":"Morace","sequence":"additional","affiliation":[{"name":"<!--label omitted: 2-->Istituto di Microbiologia, Universita\u0300 degli Studi di Milano, Milan,2 Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giovanni","family":"Fadda","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Istituto di Microbiologia, Universita\u0300Cattolica del Sacro Cuore, Rome,1 and"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1093\/clinids\/14.Supplement_1.S43"},{"key":"e_1_3_2_3_2","first-page":"338","article-title":"Detection of surgical pathogens by in vitro DNA amplification. 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