{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,6]],"date-time":"2026-02-06T03:56:07Z","timestamp":1770350167438,"version":"3.49.0"},"reference-count":27,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2004,2,1]],"date-time":"2004-02-01T00:00:00Z","timestamp":1075593600000},"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":[[2004,2]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            A conventional nested PCR and a real-time LightCycler PCR assay for detection of\n            <jats:italic>Coccidioides posadasii<\/jats:italic>\n            DNA were designed and tested in 120 clinical strains. These had been isolated from 114 patients within 10 years in Monterrey, Nuevo Leon, Mexico, known to be endemic for coccidioidomycosis. The gene encoding the specific antigen 2\/proline-rich antigen (Ag2\/PRA) was used as a target. All strains were correctly identified, whereas DNA from related members of the family\n            <jats:italic>Onygenaceae<\/jats:italic>\n            remained negative. Melting curve analysis by LightCycler and sequencing of the 526-bp product of the first PCR demonstrated either 100% identity to the GenBank sequence of the Silveira strain, now known to be\n            <jats:italic>C. posadasii<\/jats:italic>\n            (accession number\n            <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" ext-link-type=\"gen\" xlink:href=\"AF013256\">AF013256<\/jats:ext-link>\n            ), or a single silent mutation at position 1228. Length determination of two microsatellite-containing loci (GAC and 621) identified all 120 isolates as\n            <jats:italic>C. posadasii<\/jats:italic>\n            . Specific DNA was amplified by conventional nested PCR from three microscopically spherule-positive paraffin-embedded tissue samples, whereas 20 human tissue samples positive for other dimorphic fungi remained negative. Additionally, the safety of each step of a modified commercially available DNA extraction procedure was evaluated by using 10 strains. At least three steps of the protocol were demonstrated to sufficiently kill arthroconidia. This safe procedure is applicable to cultures and to clinical specimens.\n          <\/jats:p>","DOI":"10.1128\/jcm.42.2.778-783.2004","type":"journal-article","created":{"date-parts":[[2004,2,6]],"date-time":"2004-02-06T18:43:10Z","timestamp":1076092990000},"page":"778-783","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":88,"title":["PCR Assays for Identification of\n            <i>Coccidioides posadasii<\/i>\n            Based on the Nucleotide Sequence of the Antigen 2\/Proline-Rich Antigen"],"prefix":"10.1128","volume":"42","author":[{"given":"Ralf","family":"Bialek","sequence":"first","affiliation":[{"name":"Institute for Tropical Medicine, University Hospital Tu\u0308bingen"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jan","family":"Kern","sequence":"additional","affiliation":[{"name":"Institute for Tropical Medicine, University Hospital Tu\u0308bingen"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tanja","family":"Herrmann","sequence":"additional","affiliation":[{"name":"Institute for Tropical Medicine, University Hospital Tu\u0308bingen"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rolando","family":"Tijerina","sequence":"additional","affiliation":[{"name":"Departamento de Microbiologi\u0301a"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luis","family":"Cecen\u0303as","sequence":"additional","affiliation":[{"name":"Departamento de Patologi\u0301a, Facultad de Medicina, Universidad Auto\u0301noma de Nuevo Leo\u0301n, Monterrey, Nuevo Leo\u0301n, Me\u0301xico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Udo","family":"Reischl","sequence":"additional","affiliation":[{"name":"Institute of Medical Microbiology, University of Regensburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gloria M.","family":"Gonza\u0301lez","sequence":"additional","affiliation":[{"name":"Departamento de Microbiologi\u0301a"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"crossref","unstructured":"Anonymous. 2001. 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