{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,11]],"date-time":"2025-11-11T12:39:17Z","timestamp":1762864757733},"reference-count":23,"publisher":"American Society for Microbiology","issue":"5","license":[{"start":{"date-parts":[[2004,5,1]],"date-time":"2004-05-01T00:00:00Z","timestamp":1083369600000},"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":[[2004,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            We developed a rapid, sensitive, and reproducible assay to quantify\n            <jats:italic>Candida albicans ACT1<\/jats:italic>\n            ,\n            <jats:italic>CDR1<\/jats:italic>\n            ,\n            <jats:italic>CDR2<\/jats:italic>\n            ,\n            <jats:italic>ERG11<\/jats:italic>\n            , and\n            <jats:italic>MDR1<\/jats:italic>\n            mRNA using a two-step reverse transcription and LightCycler real-time PCR (RT-LightCycler PCR) method with sequence-specific hybridization probes. We compared RT-LightCycler PCR with Northern hybridization for quantitative analysis of gene expression in isolates with various fluconazole susceptibilities. Specificity of each LightCycler PCR was verified by LightCycler melting curve analysis and agarose gel electrophoresis of amplified products. Correlation of quantification results between RT-LightCycler PCR and Northern hybridization yielded correlation coefficients of \u22650.91 for all genes except\n            <jats:italic>MDR1<\/jats:italic>\n            (0.74). In this case, reduced correlation was due to the inability of Northern hybridization to accurately quantify the high\n            <jats:italic>MDR1<\/jats:italic>\n            expression in a susceptible dose-dependent isolate which was shown by RT-LightCycler PCR to overexpress\n            <jats:italic>MDR1<\/jats:italic>\n            &gt;200-fold relative to the other isolates tested. In four isolates, low levels of\n            <jats:italic>CDR2<\/jats:italic>\n            mRNA were detected by RT-LightCycler PCR but were undetectable by Northern hybridization. mRNA quantification by RT-LightCycler PCR correlates with Northern hybridization and offers additional advantages, including increased sensitivity and speed of analysis, along with lower RNA concentration requirements and an increased dynamic range of signal detection.\n          <\/jats:p>","DOI":"10.1128\/jcm.42.5.2085-2093.2004","type":"journal-article","created":{"date-parts":[[2004,5,6]],"date-time":"2004-05-06T17:29:39Z","timestamp":1083864579000},"page":"2085-2093","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":26,"title":["Rapid Quantification of Drug Resistance Gene Expression in\n            <i>Candida albicans<\/i>\n            by Reverse Transcriptase LightCycler PCR and Fluorescent Probe Hybridization"],"prefix":"10.1128","volume":"42","author":[{"given":"Joao P.","family":"Frade","sequence":"first","affiliation":[{"name":"Department of Microbiology, Faculty of Pharmacy, University of Lisbon, Lisbon, Portugal"},{"name":"Mycotic Diseases Branch, Division of Bacterial and Mycotic Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333"}]},{"given":"David W.","family":"Warnock","sequence":"additional","affiliation":[{"name":"Mycotic Diseases Branch, Division of Bacterial and Mycotic Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333"}]},{"given":"Beth A.","family":"Arthington-Skaggs","sequence":"additional","affiliation":[{"name":"Mycotic Diseases Branch, Division of Bacterial and Mycotic Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.40.12.2835"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.44.8.2081-2085.2000"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.2144\/99261rv01"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1006\/mcpr.2002.0405"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.44.10.2693-2700.2000"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.43.8.1968"},{"key":"e_1_3_2_8_2","first-page":"1026","volume":"11","year":"1993","unstructured":"Higuchi, R., C. Fockler, G. Dollinger, and R. Watson. 1993. Kinetic PCR analysis: real-time monitoring of DNA amplification reactions. Bio\/Technology11:1026-1030.","journal-title":"Bio\/Technology"},{"key":"e_1_3_2_9_2","first-page":"3171","volume":"59","year":"1999","unstructured":"Kreuzer, K. A., U. Lass, A. Bohn, O. Landt, and C. A. Schmidt. 1999. LightCycler technology for the quantitation of bcr\/abl fusion transcripts. Cancer Res.59:3171-3174.","journal-title":"Cancer Res."},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1093\/clinchem\/46.5.631"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.38.2.586-590.2000"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.42.11.2932"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.42.10.2584"},{"key":"e_1_3_2_14_2","unstructured":"National Committee for Clinical Laboratory Standards. 1997. Reference method for broth dilution antifungal susceptibility testing of yeasts. Approved standard M27-A. National Committee for Clinical Laboratory Standards Wayne Pa."},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1523-1747.2000.00845.x"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.39.10.3491-3494.2001"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1086\/344058"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.39.11.2378"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(02)00181-0"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.40.11.4308-4312.2002"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V95.3.863.003k12_863_869"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.2144\/96212rr01"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.11.2.382"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.2144\/97221bi01"}],"container-title":["Journal of Clinical Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JCM.42.5.2085-2093.2004","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JCM.42.5.2085-2093.2004","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,3,5]],"date-time":"2022-03-05T03:09:09Z","timestamp":1646449749000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/JCM.42.5.2085-2093.2004"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2004,5]]},"references-count":23,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2004,5]]}},"alternative-id":["10.1128\/JCM.42.5.2085-2093.2004"],"URL":"https:\/\/doi.org\/10.1128\/jcm.42.5.2085-2093.2004","relation":{},"ISSN":["0095-1137","1098-660X"],"issn-type":[{"value":"0095-1137","type":"print"},{"value":"1098-660X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2004,5]]},"assertion":[{"value":"2003-11-11","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2004-01-26","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2004-05-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}