{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T15:07:49Z","timestamp":1791990469283,"version":"4.4.0"},"reference-count":33,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2007,8,1]],"date-time":"2007-08-01T00:00:00Z","timestamp":1185926400000},"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":[[2007,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            The accurate diagnosis of pneumococcal disease has frequently been hampered not only by the difficulties in obtaining isolates of the organism from patient specimens but also by the misidentification of pneumococcus-like viridans group streptococci (P-LVS) as\n            <jats:italic>Streptococcus pneumoniae<\/jats:italic>\n            . This is especially critical when the specimen comes from the respiratory tract. In this study, three novel real-time PCR assays designed for the detection of specific sequence regions of the\n            <jats:italic>lytA<\/jats:italic>\n            ,\n            <jats:italic>ply<\/jats:italic>\n            , and\n            <jats:italic>psaA<\/jats:italic>\n            genes were developed (\n            <jats:italic>lytA<\/jats:italic>\n            -CDC,\n            <jats:italic>ply<\/jats:italic>\n            -CDC, and\n            <jats:italic>psaA<\/jats:italic>\n            , respectively). These assays showed high sensitivity (&lt;10 copies for\n            <jats:italic>lytA<\/jats:italic>\n            -CDC and\n            <jats:italic>ply<\/jats:italic>\n            -CDC and an approximately twofold less sensitivity for\n            <jats:italic>psaA<\/jats:italic>\n            ). Two additional real-time PCR assays for\n            <jats:italic>lytA<\/jats:italic>\n            and\n            <jats:italic>ply<\/jats:italic>\n            described previously for pneumococcal DNA detection were also evaluated. A panel of isolates consisting of 67\n            <jats:italic>S. pneumoniae<\/jats:italic>\n            isolates (44 different serotypes and 3 nonencapsulated\n            <jats:italic>S. pneumoniae<\/jats:italic>\n            isolates from conjunctivitis outbreaks) and 104 nonpneumococcal isolates was used. The 67\n            <jats:italic>S. pneumoniae<\/jats:italic>\n            isolates were reactive in all five assays. The new real-time detection assays targeting the\n            <jats:italic>lytA<\/jats:italic>\n            and\n            <jats:italic>psaA<\/jats:italic>\n            genes were the most specific for the detection of isolates confirmed to be\n            <jats:italic>S. pneumoniae<\/jats:italic>\n            , with\n            <jats:italic>lytA<\/jats:italic>\n            -CDC showing the greatest specificity. Both\n            <jats:italic>ply<\/jats:italic>\n            PCRs were positive for all isolates of\n            <jats:italic>S. pseudopneumoniae<\/jats:italic>\n            , along with 13 other isolates of other P-LVS isolates confirmed to be non-\n            <jats:italic>S. pneumoniae<\/jats:italic>\n            by DNA-DNA reassociation. Thus, the use of the\n            <jats:italic>ply<\/jats:italic>\n            gene for the detection of pneumococci can lead to false-positive reactions in the presence of P-LVS. The five assays were applied to 15 culture-positive cerebrospinal fluid specimens with 100% sensitivity; and serum and ear fluid specimens were also evaluated. Both the\n            <jats:italic>lytA<\/jats:italic>\n            -CDC and\n            <jats:italic>psaA<\/jats:italic>\n            assays, particularly the\n            <jats:italic>lytA<\/jats:italic>\n            -CDC assay, have improved specificities compared with those of currently available assays and should therefore be considered the assays of choice for the detection of pneumococcal DNA, particularly when upper respiratory P-LVS might be present in the clinical specimen.\n          <\/jats:p>","DOI":"10.1128\/jcm.02498-06","type":"journal-article","created":{"date-parts":[[2007,5,31]],"date-time":"2007-05-31T00:54:28Z","timestamp":1180572868000},"page":"2460-2466","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":556,"title":["Evaluation and Improvement of Real-Time PCR Assays Targeting\n            <i>lytA<\/i>\n            ,\n            <i>ply<\/i>\n            , and\n            <i>psaA<\/i>\n            Genes for Detection of Pneumococcal DNA"],"prefix":"10.1128","volume":"45","author":[{"given":"Maria da Gloria S.","family":"Carvalho","sequence":"first","affiliation":[{"name":"Division of Bacterial Diseases"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria Lucia","family":"Tondella","sequence":"additional","affiliation":[{"name":"Division of Bacterial Diseases"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karen","family":"McCaustland","sequence":"additional","affiliation":[{"name":"Division of Scientific Resources, Centers for Disease Control and Prevention"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luciana","family":"Weidlich","sequence":"additional","affiliation":[{"name":"Centro de Desenvolvimento Cienti\u0301fico e Tecnolo\u0301gico, Fundac\u0327a\u0303o Estadual de Produc\u0327a\u0303o e Pesquisa em Sau\u0301de, Porto Alegre, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lesley","family":"McGee","sequence":"additional","affiliation":[{"name":"Hubert Department of Global Health, Emory University, Atlanta, Georgia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leonard W.","family":"Mayer","sequence":"additional","affiliation":[{"name":"Division of Bacterial Diseases"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arnold","family":"Steigerwalt","sequence":"additional","affiliation":[{"name":"Division of Bacterial Diseases"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melissa","family":"Whaley","sequence":"additional","affiliation":[{"name":"Division of Bacterial Diseases"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard R.","family":"Facklam","sequence":"additional","affiliation":[{"name":"Division of Bacterial Diseases"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barry","family":"Fields","sequence":"additional","affiliation":[{"name":"Division of Bacterial Diseases"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"George","family":"Carlone","sequence":"additional","affiliation":[{"name":"Division of Bacterial Diseases"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edwin W.","family":"Ades","sequence":"additional","affiliation":[{"name":"Division of Bacterial Diseases"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ron","family":"Dagan","sequence":"additional","affiliation":[{"name":"Pediatric Infectious Diseases Unit, Soroka University Medical Center, Beer Sheva, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacquelyn S.","family":"Sampson","sequence":"additional","affiliation":[{"name":"Division of Bacterial Diseases"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","first-page":"1","volume":"49","year":"2000","unstructured":"Advisory Committee on Immunization Practices. 2000. 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