{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,18]],"date-time":"2026-10-18T03:13:58Z","timestamp":1792293238350,"version":"4.4.0"},"update-to":[{"DOI":"10.1371\/journal.pntd.0009765","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2021,10,1]],"date-time":"2021-10-01T00:00:00Z","timestamp":1633046400000}}],"reference-count":26,"publisher":"Public Library of Science (PLoS)","issue":"9","license":[{"start":{"date-parts":[[2021,9,16]],"date-time":"2021-09-16T00:00:00Z","timestamp":1631750400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/publicdomain\/zero\/1.0\/"}],"funder":[{"DOI":"10.13039\/100004857","name":"Wadsworth Center","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100004857","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100004856","name":"New York State Department of Health","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100004856","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000030","name":"Centers for Disease Control and Prevention","doi-asserted-by":"publisher","award":["NU50CK000516"],"award-info":[{"award-number":["NU50CK000516"]}],"id":[{"id":"10.13039\/100000030","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosntds.org"],"crossmark-restriction":false},"short-container-title":["PLoS Negl Trop Dis"],"abstract":"<jats:p>\n                    Coccidioidomycosis (Valley fever) is a pulmonary and systemic fungal disease with increasing incidence and expanding endemic areas. The differentiation of etiologic agents\n                    <jats:italic>Coccidioides immitis<\/jats:italic>\n                    and\n                    <jats:italic>C<\/jats:italic>\n                    .\n                    <jats:italic>posadasii<\/jats:italic>\n                    remains problematic in the clinical laboratories as conventional PCR and satellite typing schemes are not facile. Therefore, we developed Cy5- and FAM-labeled TaqMan-probes for duplex real-time PCR assay for rapid differentiation of\n                    <jats:italic>C<\/jats:italic>\n                    .\n                    <jats:italic>immitis<\/jats:italic>\n                    and\n                    <jats:italic>C<\/jats:italic>\n                    .\n                    <jats:italic>posadasii<\/jats:italic>\n                    from culture and clinical specimens. The\n                    <jats:italic>RRA2<\/jats:italic>\n                    gene encoding proline-rich antigen 2, specific for\n                    <jats:italic>Coccidioides<\/jats:italic>\n                    genus, was the source for the first set of primers and probe.\n                    <jats:italic>Coccidioides immitis<\/jats:italic>\n                    contig 2.2 (GenBank: AAEC02000002.1) was used to design the second set of primers and probe. The second primers\/probe did not amplify the corresponding\n                    <jats:italic>C<\/jats:italic>\n                    .\n                    <jats:italic>posadasii<\/jats:italic>\n                    DNA, because of an 86-bp deletion in the contig. The assay was highly sensitive with limit of detection of 0.1 pg gDNA\/PCR reaction, which was equivalent to approximately ten genome copies of\n                    <jats:italic>C<\/jats:italic>\n                    .\n                    <jats:italic>immitis<\/jats:italic>\n                    or\n                    <jats:italic>C<\/jats:italic>\n                    .\n                    <jats:italic>posadasii<\/jats:italic>\n                    . The assay was highly specific with no cross-reactivity to the wide range of fungal and bacterial pathogens. Retrospective analysis of fungal isolates and primary specimens submitted from 1995 to 2020 confirmed 168 isolates and four primary specimens as\n                    <jats:italic>C<\/jats:italic>\n                    .\n                    <jats:italic>posadasii<\/jats:italic>\n                    and 30 isolates as\n                    <jats:italic>C<\/jats:italic>\n                    .\n                    <jats:italic>immitis<\/jats:italic>\n                    from human coccidioidomycosis cases, while all eight primary samples from two animals (rhesus monkey and rhinoceros) were confirmed as\n                    <jats:italic>C<\/jats:italic>\n                    .\n                    <jats:italic>posadasii<\/jats:italic>\n                    . A preliminary analysis of cerebrospinal fluid (CSF) and pleural fluid samples showed positive correlation between serology tests and real-time PCR for two of the 15 samples. The\n                    <jats:italic>Coccidioides<\/jats:italic>\n                    spp. duplex real-time PCR will allow rapid differentiation of\n                    <jats:italic>C<\/jats:italic>\n                    .\n                    <jats:italic>immitis<\/jats:italic>\n                    and\n                    <jats:italic>C<\/jats:italic>\n                    .\n                    <jats:italic>posadasii<\/jats:italic>\n                    from clinical specimens and further augment the treatment and surveillance of coccidioidomycosis.\n                  <\/jats:p>","DOI":"10.1371\/journal.pntd.0009765","type":"journal-article","created":{"date-parts":[[2021,9,16]],"date-time":"2021-09-16T13:44:15Z","timestamp":1631799855000},"page":"e0009765","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":13,"title":["Real-time PCR assay for detection and differentiation of Coccidioides immitis and Coccidioides 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L.","family":"Crucillo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3922-9676","authenticated-orcid":true,"given":"Vishnu","family":"Chaturvedi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2021,9,16]]},"reference":[{"issue":"9","key":"pntd.0009765.ref001","doi-asserted-by":"crossref","first-page":"1217","DOI":"10.1086\/496991","article-title":"Coccidioidomycosis","volume":"41","author":"JN Galgiani","year":"2005","journal-title":"Clin Infect Dis"},{"issue":"6","key":"pntd.0009765.ref002","doi-asserted-by":"crossref","first-page":"847","DOI":"10.1093\/cid\/cis1028","article-title":"Coccidioidomycosis acquired in Washington State","volume":"56","author":"N Marsden-Haug","year":"2013","journal-title":"Clin Infect Dis"},{"issue":"20","key":"pntd.0009765.ref003","first-page":"450","article-title":"Notes 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