{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,4]],"date-time":"2026-03-04T16:55:36Z","timestamp":1772643336836,"version":"3.50.1"},"reference-count":37,"publisher":"American Society for Microbiology","issue":"5","license":[{"start":{"date-parts":[[2011,5,1]],"date-time":"2011-05-01T00:00:00Z","timestamp":1304208000000},"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":[[2011,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            <jats:named-content content-type=\"genus-species\">Pneumocystis jirovecii<\/jats:named-content>\n            pneumonia (PcP) is a major cause of respiratory illness in patients with AIDS. The identification of multiple single-nucleotide polymorphisms (SNPs) at three distinct\n            <jats:named-content content-type=\"genus-species\">P. jirovecii<\/jats:named-content>\n            loci encoding dihydrofolate reductase (\n            <jats:italic>DHFR<\/jats:italic>\n            ), mitochondrial large-subunit rRNA (\n            <jats:italic>mtLSU rRNA<\/jats:italic>\n            ), and superoxide dismutase (\n            <jats:italic>SOD<\/jats:italic>\n            ) was achieved using multiplex-PCR (MPCR) followed by direct sequencing and two single-base extension (SBE) techniques. Four SNPs (\n            <jats:italic>DHFR312<\/jats:italic>\n            ,\n            <jats:italic>mt85<\/jats:italic>\n            ,\n            <jats:italic>SOD215<\/jats:italic>\n            , and\n            <jats:italic>SOD110<\/jats:italic>\n            ), correlated previously with parameters of disease, were amplified and genotyped simultaneously. The concordance of results between the standard sequencing technique (direct sequencing) and SBE analysis was 96.9% for the acrylamide gel electrophoresis and 98.4% for the capillary electrophoresis. The cross-genetic analysis established several statistical associations among the SNPs studied:\n            <jats:italic>mt85C-SOD110T<\/jats:italic>\n            ,\n            <jats:italic>SOD110T-SOD215C<\/jats:italic>\n            , and\n            <jats:italic>SOD110C-SOD215T<\/jats:italic>\n            . These results were confirmed by cluster analysis. Data showed that among the isolates with low to moderate parasite burden, the highest percentages of\n            <jats:italic>DHFR312C<\/jats:italic>\n            ,\n            <jats:italic>mt85C<\/jats:italic>\n            ,\n            <jats:italic>SOD110T<\/jats:italic>\n            , and\n            <jats:italic>SOD215C<\/jats:italic>\n            were detected, whereas for high parasite burden cases the highest frequencies were observed among isolates with\n            <jats:italic>DHFR312T<\/jats:italic>\n            ,\n            <jats:italic>mt85T<\/jats:italic>\n            ,\n            <jats:italic>SOD110C<\/jats:italic>\n            , and\n            <jats:italic>SOD215T<\/jats:italic>\n            . The polymorphisms studied were shown to be suitable genetic targets potentially correlated with PcP clinical data that can be used as predictors of outcome in further studies to help clinical decision-making in the management of PcP. The MPCR\/SBE protocol described for the first time in the present study was shown to be a rapid, highly accurate method for genotyping\n            <jats:named-content content-type=\"genus-species\">P. jirovecii<\/jats:named-content>\n            SNPs encoded by different loci that could be used for epidemiological studies and as an additional procedure for the prognostic classification and diagnosis of PcP.\n          <\/jats:p>","DOI":"10.1128\/jcm.02303-10","type":"journal-article","created":{"date-parts":[[2011,3,10]],"date-time":"2011-03-10T03:53:13Z","timestamp":1299729193000},"page":"1810-1815","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":37,"title":["Clinical Relevance of Multiple Single-Nucleotide Polymorphisms in Pneumocystis jirovecii Pneumonia: Development of a Multiplex PCR-Single-Base-Extension Methodology"],"prefix":"10.1128","volume":"49","author":[{"given":"F.","family":"Esteves","sequence":"first","affiliation":[{"name":"Unidade de Protozo\u00e1rios Oportunistas\/VIH e Outras Protozooses\u2013CMDT, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Lisbon, Portugal"}]},{"given":"J.","family":"Gaspar","sequence":"additional","affiliation":[{"name":"Departamento de Gen\u00e9tica, Faculdade de Ci\u00eancias M\u00e9dicas, Universidade Nova de Lisboa, Lisbon, Portugal"}]},{"given":"B.","family":"De Sousa","sequence":"additional","affiliation":[{"name":"Unidade de Epidemiologia e Bioestat\u00edstica\u2013CMDT, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Lisbon, Portugal"}]},{"given":"F.","family":"Antunes","sequence":"additional","affiliation":[{"name":"Cl\u00ednica das Doen\u00e7as Infecciosas, Hospital de Santa Maria, Lisbon, Portugal"}]},{"given":"K.","family":"Mansinho","sequence":"additional","affiliation":[{"name":"Servi\u00e7o de Doen\u00e7as Infecciosas, Hospital de Egas Moniz, Lisbon, Portugal"}]},{"given":"O.","family":"Matos","sequence":"additional","affiliation":[{"name":"Unidade de Protozo\u00e1rios Oportunistas\/VIH e Outras Protozooses\u2013CMDT, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Lisbon, 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