{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T02:13:44Z","timestamp":1792116824365,"version":"4.4.0"},"reference-count":29,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2007,12,17]],"date-time":"2007-12-17T00:00:00Z","timestamp":1197849600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"Ministry of Science and Technology of China","award":["2003BA712A11-20"],"award-info":[{"award-number":["2003BA712A11-20"]}]},{"name":"Bureau of Science and Technology of Beijing","award":["Y0204004040131"],"award-info":[{"award-number":["Y0204004040131"]}]},{"name":"Young Award of Bureau Science and Technology of Beijing","award":["2004B02"],"award-info":[{"award-number":["2004B02"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pediatric Pulmonology"],"published-print":{"date-parts":[[2008,2]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Objectives<\/jats:title><jats:p>To develop and evaluate a novel method for simultaneous identification of 12 potential bacterial pathogens in children with community\u2010acquired pneumonia.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>A multiplex PCR\u2010based reverse line blot (mPCR\/RLB) assay was developed, to identify 12 respiratory bacterial pathogens, namely <jats:italic>Staphylococcus aureus<\/jats:italic>, <jats:italic>Streptococcus pneumoniae<\/jats:italic>, <jats:italic>Streptococcus pyogenes<\/jats:italic>, <jats:italic>Moraxella catarrhalis<\/jats:italic>, <jats:italic>Haemophilus influenzae<\/jats:italic>, <jats:italic>Haemophilus influenzae<\/jats:italic> type b, <jats:italic>Bordetella pertussis<\/jats:italic>, <jats:italic>Klebsiella pneumoniae<\/jats:italic>, <jats:italic>Legionella pneumophila<\/jats:italic>, <jats:italic>Mycobacterium tuberculosis<\/jats:italic>, <jats:italic>Chlamydia pneumoniae<\/jats:italic>, <jats:italic>Mycoplasma pneumoniae<\/jats:italic>, and single (uniplex) PCRs were used for <jats:italic>S. pneumoniae<\/jats:italic> and <jats:italic>H. influenzae<\/jats:italic> only. In a preliminary evaluation, we compared the results of mPCR\/RLB with those of single (uniplex) PCRs and culture of nasopharyngeal aspirates (NPAs) from 100 children under 5 years, admitted to Beijing Children's Hospital between October 2004 and May 2005, with pneumonia.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Reference strains and clinical isolates of all 12 target species were correctly identified by mPCR\/RLB. Potential pathogens were isolated from one blood culture and 26% of respiratory cultures. One or more pathogens were identified in 70% of respiratory specimens\u2014by mPCR\/RLB in 63%, uPCR only in another 3%, culture only in 2%, and culture plus uPCR in 2%. The species most commonly identified were <jats:italic>S. pneumoniae<\/jats:italic> (54%) and <jats:italic>H. influenzae<\/jats:italic> (38%, including type b, 4%). Cultures were not performed for <jats:italic>B. pertussis, M. tuberculosis<\/jats:italic>, <jats:italic>C. pneumoniae<\/jats:italic> or <jats:italic>M<\/jats:italic>. <jats:italic>pneumoniae<\/jats:italic> but each was identified by mPCR\/RLB in between one and four specimens. Two or more potential pathogens were identified in 35% of specimens. Ten of 14 <jats:italic>S. pneumoniae<\/jats:italic> isolates belonged to serotypes represented in the 11\u2010valent pneumococcal conjugate vaccine.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>The mPCR\/RLB assay is a sensitive tool for identification of respiratory pathogens, including mixed infections and bacteria requiring special culture methods. Pediatr Pulmonol. 2008; 43:150\u2013159. \u00a9 2007 Wiley\u2010Liss, Inc.<\/jats:p><\/jats:sec>","DOI":"10.1002\/ppul.20749","type":"journal-article","created":{"date-parts":[[2007,12,18]],"date-time":"2007-12-18T00:16:13Z","timestamp":1197936973000},"page":"150-159","source":"Crossref","is-referenced-by-count":38,"title":["A multiplex PCR\u2010based reverse line blot hybridization (mPCR\/RLB) assay for detection of bacterial respiratory pathogens in children with pneumonia"],"prefix":"10.1002","volume":"43","author":[{"given":"Yajuan","family":"Wang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fanrong","family":"Kong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yonghong","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gwendolyn L.","family":"Gilbert","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2007,12,26]]},"reference":[{"key":"e_1_2_1_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(05)71877-8"},{"key":"e_1_2_1_3_2","first-page":"1159","article-title":"Fever in respiratory virus infections","volume":"140","author":"Putto A","year":"1986","journal-title":"Am J Dis Child"},{"key":"e_1_2_1_4_2","first-page":"899","article-title":"Community\u2010acquired pneumonia in infants and children","volume":"70","author":"Ostapchuk M","year":"2004","journal-title":"Am Fam Physician"},{"key":"e_1_2_1_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.44.5.1887-1891.2006"},{"key":"e_1_2_1_6_2","doi-asserted-by":"publisher","DOI":"10.1099\/jmm.0.46244-0"},{"key":"e_1_2_1_7_2","doi-asserted-by":"publisher","DOI":"10.1097\/00006454-200101000-00010"},{"key":"e_1_2_1_8_2","doi-asserted-by":"publisher","DOI":"10.1001\/jama.279.4.308"},{"key":"e_1_2_1_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2006.404"},{"key":"e_1_2_1_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.diagmicrobio.2004.08.014"},{"key":"e_1_2_1_11_2","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/171.2.479"},{"key":"e_1_2_1_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.42.9.4268-4274.2004"},{"key":"e_1_2_1_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.41.4.1386-1390.2003"},{"key":"e_1_2_1_14_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.42.8.3813-3815.2004"},{"key":"e_1_2_1_15_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.43.2.565-571.2005"},{"key":"e_1_2_1_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.41.7.3327-3330.2003"},{"key":"e_1_2_1_17_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.41.10.4565-4572.2003"},{"key":"e_1_2_1_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.42.7.3219-3224.2004"},{"key":"e_1_2_1_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.70.3.1483-1486.2004"},{"key":"e_1_2_1_20_2","doi-asserted-by":"publisher","DOI":"10.1086\/319213"},{"key":"e_1_2_1_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/0732-8893(95)00175-1"},{"key":"e_1_2_1_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1521-6616(03)00042-1"},{"key":"e_1_2_1_23_2","doi-asserted-by":"publisher","DOI":"10.1007\/s004310100775"},{"key":"e_1_2_1_24_2","doi-asserted-by":"publisher","DOI":"10.1097\/00006454-200003000-00003"},{"key":"e_1_2_1_25_2","doi-asserted-by":"publisher","DOI":"10.1097\/00006454-200209000-00005"},{"key":"e_1_2_1_26_2","doi-asserted-by":"publisher","DOI":"10.1097\/00006454-200204000-00002"},{"key":"e_1_2_1_27_2","doi-asserted-by":"publisher","DOI":"10.1097\/00006454-200004000-00006"},{"key":"e_1_2_1_28_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1651-2227.1993.tb12624.x"},{"key":"e_1_2_1_29_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF02072512"},{"key":"e_1_2_1_30_2","doi-asserted-by":"publisher","DOI":"10.1097\/01.inf.0000078373.54515.40"}],"container-title":["Pediatric Pulmonology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fppul.20749","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fppul.20749","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/ppul.20749","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,11,14]],"date-time":"2023-11-14T16:50:19Z","timestamp":1699980619000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/ppul.20749"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2007,12,26]]},"references-count":29,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2008,2]]}},"alternative-id":["10.1002\/ppul.20749"],"URL":"https:\/\/doi.org\/10.1002\/ppul.20749","archive":["Portico"],"relation":{},"ISSN":["8755-6863","1099-0496"],"issn-type":[{"value":"8755-6863","type":"print"},{"value":"1099-0496","type":"electronic"}],"subject":[],"published":{"date-parts":[[2007,12,26]]}}}