{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T03:54:55Z","timestamp":1792641295567,"version":"4.5.0"},"reference-count":61,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[2014,4,1]],"date-time":"2014-04-01T00:00:00Z","timestamp":1396310400000},"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":[[2014,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            A molecular diagnostic technique based on real-time PCR was developed for the simultaneous detection of three of the most frequent causative agents of fungal opportunistic pneumonia in AIDS patients:\n            <jats:named-content content-type=\"genus-species\">Pneumocystis jirovecii<\/jats:named-content>\n            ,\n            <jats:named-content content-type=\"genus-species\">Histoplasma capsulatum<\/jats:named-content>\n            , and\n            <jats:named-content content-type=\"genus-species\">Cryptococcus neoformans<\/jats:named-content>\n            \/\n            <jats:named-content content-type=\"genus-species\">Cryptococcus gattii<\/jats:named-content>\n            . This technique was tested in cultured strains and in clinical samples from HIV-positive patients. The methodology used involved species-specific molecular beacon probes targeted to the internal transcribed spacer regions of the rDNA. An internal control was also included in each assay. The multiplex real-time PCR assay was tested in 24 clinical strains and 43 clinical samples from AIDS patients with proven fungal infection. The technique developed showed high reproducibility (\n            <jats:italic>r<\/jats:italic>\n            <jats:sup>2<\/jats:sup>\n            of &gt;0.98) and specificity (100%). For\n            <jats:named-content content-type=\"genus-species\">H. capsulatum<\/jats:named-content>\n            and\n            <jats:named-content content-type=\"genus-species\">Cryptococcus<\/jats:named-content>\n            spp., the detection limits of the method were 20 and 2 fg of genomic DNA\/20 \u03bcl reaction mixture, respectively, while for\n            <jats:named-content content-type=\"genus-species\">P. jirovecii<\/jats:named-content>\n            the detection limit was 2.92 log\n            <jats:sub>10<\/jats:sub>\n            copies\/20 \u03bcl reaction mixture. The sensitivity\n            <jats:italic>in vitro<\/jats:italic>\n            was 100% for clinical strains and 90.7% for clinical samples. The assay was positive for 92.5% of the patients. For one of the patients with proven histoplasmosis,\n            <jats:named-content content-type=\"genus-species\">P. jirovecii<\/jats:named-content>\n            was also detected in a bronchoalveolar lavage sample. No PCR inhibition was detected. This multiplex real-time PCR technique is fast, sensitive, and specific and may have clinical applications.\n          <\/jats:p>","DOI":"10.1128\/jcm.02895-13","type":"journal-article","created":{"date-parts":[[2014,1,30]],"date-time":"2014-01-30T04:55:23Z","timestamp":1391057723000},"page":"1168-1176","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":65,"title":["A Multiplex Real-Time PCR Assay for Identification of Pneumocystis jirovecii, Histoplasma capsulatum, and Cryptococcus neoformans\/Cryptococcus gattii in Samples from AIDS Patients with Opportunistic Pneumonia"],"prefix":"10.1128","volume":"52","author":[{"given":"Sara","family":"Gago","sequence":"first","affiliation":[{"name":"Mycology Department, National Centre for Microbiology, Instituto de Salud Carlos III, Majadahonda, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristina","family":"Esteban","sequence":"additional","affiliation":[{"name":"Department of Clinical Microbiology, Hospital Clinic de Barcelona, Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Clara","family":"Valero","sequence":"additional","affiliation":[{"name":"Mycology Department, National Centre for Microbiology, Instituto de Salud Carlos III, Majadahonda, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00d3scar","family":"Zaragoza","sequence":"additional","affiliation":[{"name":"Mycology Department, National Centre for Microbiology, Instituto de Salud Carlos III, Majadahonda, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jorge","family":"Puig de la Bellacasa","sequence":"additional","affiliation":[{"name":"Department of Clinical Microbiology, Hospital Clinic de Barcelona, Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mar\u00eda Jos\u00e9","family":"Buitrago","sequence":"additional","affiliation":[{"name":"Mycology Department, National Centre for Microbiology, Instituto de Salud Carlos III, Majadahonda, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1513\/pats.201009-062WR"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0058812"},{"key":"e_1_3_2_4_2","first-page":"263","article-title":"Pulmonary complications of HIV infection","volume":"59","author":"Rosen MJ","year":"1992","unstructured":"RosenMJ. 1992. 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