{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,26]],"date-time":"2026-03-26T22:16:18Z","timestamp":1774563378285,"version":"3.50.1"},"reference-count":24,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2003,2,1]],"date-time":"2003-02-01T00:00:00Z","timestamp":1044057600000},"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":[[2003,2]]},"abstract":"ABSTRACT<\/jats:title>\n \n Chlamydia pneumoniae<\/jats:italic>\n , an important respiratory pathogen, is difficult to culture, and detection rates by conventional PCRs vary considerably. A new quantitative\n ompA<\/jats:italic>\n -based real-time PCR assay based on TaqMan technology for detection of\n C. pneumoniae<\/jats:italic>\n in respiratory samples is described, and its performance in terms of sensitivity and reproducibility is compared with those of four published conventional PCRs (one single-step PCR targeting a cloned\n Pst<\/jats:italic>\n I fragment; two nested PCRs, one targeting the 16S rRNA gene followed by hybridization and the other targeting the\n ompA<\/jats:italic>\n gene; and a touchdown enzyme time-release [TETR] PCR also targeting the 16S rRNA gene). Both\n ompA<\/jats:italic>\n -based PCRs showed the best analytical sensitivity. All five assays could detect even lower target levels from spiked sputum, with the 16S rRNA assays performing better than the\n ompA<\/jats:italic>\n -based nested PCR (10\n \u22126<\/jats:sup>\n inclusion-forming units [IFU] were detected in four of four and two of four replicates by the 16S rRNA TETR PCR and the 16S rRNA nested PCR, respectively). In general, the\n ompA<\/jats:italic>\n -based real-time protocol produced the most consistent positive results for all replicates tested down to 10\n \u22126<\/jats:sup>\n IFU. Eight of 45 patient sputum specimens (18%) were\n C. pneumoniae<\/jats:italic>\n DNA positive in at least one of four replicates tested by at least one assay. Without taking into consideration the analytical sensitivity or the reproducibility of the test results, the numbers of\n C. pneumoniae<\/jats:italic>\n DNA-positive sputum specimens (\n n<\/jats:italic>\n = 8) were four, three, two, two, and one for the 16S rRNA TETR assay, the\n Pst<\/jats:italic>\n I-based single-step PCR, the\n ompA<\/jats:italic>\n -based real-time PCR, the\n ompA<\/jats:italic>\n -based nested touchdown PCR, and the 16S rRNA-based nested PCR, respectively. However, the overall rate of concordance of positive results was low. Only one cell culture-positive sputum specimen was positive by four of five assays (14 of 16 replicates; mean cycle threshold value, 25; 10\n 8<\/jats:sup>\n particles\/ml of sputum). Thirty-seven specimens were\n C. pneumoniae<\/jats:italic>\n negative by all five assays for all replicates tested, as were all negative controls (\n n<\/jats:italic>\n = 65 to 100 per testing panel). No PCR inhibitors were detected by real-time PCR or by the 16S rRNA-based nested assay. We confirm that the analytical sensitivity of an assay for the detection of\n C. pneumoniae<\/jats:italic>\n does not necessarily predict its ability to detect its target in sputum. A quantitative, fast, and easy-to-handle diagnostic approach such as the\n ompA<\/jats:italic>\n -based real-time TaqMan PCR described here might improve the detection of\n C. pneumoniae<\/jats:italic>\n in respiratory samples.\n <\/jats:p>","DOI":"10.1128\/jcm.41.2.592-600.2003","type":"journal-article","created":{"date-parts":[[2003,2,6]],"date-time":"2003-02-06T20:03:31Z","timestamp":1044561811000},"page":"592-600","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":83,"title":["Comparison of a New Quantitative\n ompA<\/i>\n -Based Real-Time PCR TaqMan Assay for Detection of\n Chlamydia pneumoniae<\/i>\n DNA in Respiratory Specimens with Four Conventional PCR Assays"],"prefix":"10.1128","volume":"41","author":[{"given":"Petra","family":"Apfalter","sequence":"first","affiliation":[{"name":"Division of Clinical Microbiology, Department of Hygiene and Medical Microbiology, University of Vienna, Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wolfgang","family":"Barousch","sequence":"additional","affiliation":[{"name":"Division of Clinical Microbiology, Department of Hygiene and Medical Microbiology, University of Vienna, Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marion","family":"Nehr","sequence":"additional","affiliation":[{"name":"Division of Clinical Microbiology, Department of Hygiene and Medical Microbiology, University of Vienna, Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Athanasios","family":"Makristathis","sequence":"additional","affiliation":[{"name":"Division of Clinical Microbiology, Department of Hygiene and Medical Microbiology, University of Vienna, Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Birgit","family":"Willinger","sequence":"additional","affiliation":[{"name":"Division of Clinical Microbiology, Department of Hygiene and Medical Microbiology, University of Vienna, Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manfred","family":"Rotter","sequence":"additional","affiliation":[{"name":"Division of Clinical Microbiology, Department of Hygiene and Medical Microbiology, University of Vienna, Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexander M.","family":"Hirschl","sequence":"additional","affiliation":[{"name":"Division of Clinical Microbiology, Department of Hygiene and Medical Microbiology, University of Vienna, Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_1_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.39.2.519-524.2001"},{"key":"e_1_3_1_3_2","doi-asserted-by":"publisher","DOI":"10.1007\/s100960100554"},{"key":"e_1_3_1_4_2","doi-asserted-by":"publisher","DOI":"10.1007\/s100960050481"},{"key":"e_1_3_1_5_2","doi-asserted-by":"publisher","DOI":"10.1007\/s004300050071"},{"key":"e_1_3_1_6_2","doi-asserted-by":"publisher","DOI":"10.1086\/315609"},{"key":"e_1_3_1_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.37.12.3791-3799.1999"},{"key":"e_1_3_1_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.15.1.1-20.2002"},{"key":"e_1_3_1_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.30.2.434-439.1992"},{"key":"e_1_3_1_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.40.7.2357-2362.2002"},{"key":"e_1_3_1_11_2","doi-asserted-by":"publisher","DOI":"10.1086\/322632"},{"key":"e_1_3_1_12_2","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/168.5.1231"},{"key":"e_1_3_1_13_2","doi-asserted-by":"publisher","DOI":"10.1093\/clinids\/14.1.178"},{"key":"e_1_3_1_14_2","doi-asserted-by":"publisher","DOI":"10.1093\/clinids\/20.5.1174"},{"key":"e_1_3_1_15_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.35.3.620-623.1997"},{"key":"e_1_3_1_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.8.4.451"},{"key":"e_1_3_1_17_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.40.6.2273-2274.2002"},{"key":"e_1_3_1_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.38.3.1085-1093.2000"},{"key":"e_1_3_1_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.38.7.2622-2627.2000"},{"key":"e_1_3_1_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.35.8.2043-2046.1997"},{"key":"e_1_3_1_21_2","first-page":"153","volume":"111","year":"1999","unstructured":"Nadrchal, R., A. Makristathis, P. Apfalter, M. Rotter, W. Trubel, I. Huk, P. Polterauer, and A. M. Hirschl. 1999. Detection of Chlamydia pneumoniae DNA in atheromatous tissues by polymerase chain reaction. Wien. Klin. Wochenschr.111:153-156.","journal-title":"Wien. Klin. Wochenschr."},{"key":"e_1_3_1_22_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.30.8.1968-1971.1992"},{"key":"e_1_3_1_23_2","first-page":"1796","volume":"29","year":"2001","unstructured":"Smieja, M., J. B. Mahony, C. H. Goldsmith, S. Chong, A. Petrich, and M. Chernesky. 2001. Replicate PCR testing and probit analysis for detection and quantitation of Chlamydia pneumoniae in clinical specimens. J. Clin. Microbiol.29:1796-1801.","journal-title":"J. Clin. 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