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The method relies on the amplification of two regions within the bacterial 16S rRNA gene, using universal PCR primers and querying the identity of specific single-nucleotide polymorphisms within the amplified regions in a subsequent LDR. The ligation products vary in color and size and are separated by CE. Each organism generates a specific pattern of ligation products, which can be used to distinguish the pathogens using an automated software program we developed for that purpose. The assay has been verified on 315 clinical isolates and demonstrated a detection sensitivity of 98%. Additionally, 484 seeded blood cultures were tested, with a detection sensitivity of 97.7%. The ability to identify geographically variant strains of the organisms was determined by testing 132 isolates obtained from across the United States. In summary, the PCR-LDR-CE assay can successfully identify, in a multiplexed fashion, a panel of 20 blood-borne pathogens with high sensitivity and specificity.<\/jats:p>","DOI":"10.1128\/jcm.00226-07","type":"journal-article","created":{"date-parts":[[2007,4,12]],"date-time":"2007-04-12T00:34:25Z","timestamp":1176338065000},"page":"1927-1935","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":40,"title":["Multiplexed Identification of Blood-Borne Bacterial Pathogens by Use of a Novel 16S rRNA Gene PCR-Ligase Detection Reaction-Capillary Electrophoresis Assay"],"prefix":"10.1128","volume":"45","author":[{"given":"Maneesh R.","family":"Pingle","sequence":"first","affiliation":[{"name":"Department of Microbiology"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kathleen","family":"Granger","sequence":"additional","affiliation":[{"name":"Department of Microbiology"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philip","family":"Feinberg","sequence":"additional","affiliation":[{"name":"Department of Microbiology"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rebecca","family":"Shatsky","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bram","family":"Sterling","sequence":"additional","affiliation":[{"name":"Department of Microbiology"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark","family":"Rundell","sequence":"additional","affiliation":[{"name":"Department of Microbiology"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eric","family":"Spitzer","sequence":"additional","affiliation":[{"name":"Department of Pathology, Stony Brook University Medical Center, Stony Brook, New York"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Davise","family":"Larone","sequence":"additional","affiliation":[{"name":"Department of Microbiology"},{"name":"Department of Pathology and Laboratory Medicine"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Linnie","family":"Golightly","sequence":"additional","affiliation":[{"name":"Department of Microbiology"},{"name":"Division of Infectious Disease and International Medicine, Weill Medical College of Cornell University, New York, New York"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francis","family":"Barany","sequence":"additional","affiliation":[{"name":"Department of Microbiology"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.38.2.781-788.2000"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.88.1.189"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1101\/gr.1.1.5"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.37.5.1415-1418.1999"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1369-5274(02)00362-4"},{"key":"e_1_3_2_7_2","first-page":"1258","volume":"36","year":"1990","unstructured":"Bottger, E. 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