{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,22]],"date-time":"2025-10-22T10:36:19Z","timestamp":1761129379572,"version":"build-2065373602"},"reference-count":12,"publisher":"Wiley","issue":"23-24","license":[{"start":{"date-parts":[[2016,9,27]],"date-time":"2016-09-27T00:00:00Z","timestamp":1474934400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100003710","name":"Korea Health Industry Development Institute","doi-asserted-by":"publisher","award":["H14C2658"],"award-info":[{"award-number":["H14C2658"]}],"id":[{"id":"10.13039\/501100003710","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Electrophoresis"],"published-print":{"date-parts":[[2016,12]]},"abstract":"<jats:p>Early detection of pathogens from blood and identification of their drug resistance are essential for sepsis management. However, conventional culture\u2010based methods require relatively longer time to identify drug\u2010resistant pathogens, which delays therapeutic decisions. For precise multiplex detection of drug\u2010resistant Gram\u2010positive pathogens, we developed a method by using stuffer\u2010free multiplex ligation\u2010dependent probe amplification (MLPA) coupled with high\u2010resolution CE single\u2010strand conformation polymorphisms (CE\u2010SSCP) system. We designed three probe sets for genes specific to Gram\u2010positive species (<jats:italic>Staphylococcus aureus<\/jats:italic>: <jats:italic>nuc<\/jats:italic>, <jats:italic>Enterococcus faecium<\/jats:italic>: <jats:italic>sodA<\/jats:italic>, and <jats:italic>Streptococcus pneumoniae<\/jats:italic>: <jats:italic>lytA<\/jats:italic>) and two sets for genes associated with drug resistance (<jats:italic>mecA<\/jats:italic> and <jats:italic>vanA<\/jats:italic>) to discriminate major Gram\u2010positive pathogens with the resistance. A total of 94 different strains (34 reference strains and 60 clinical isolates) were used to validate this method and strain\u2010specific peaks were successfully observed for all the strains. To improve sensitivity of the method, a target\u2010specific preamplification step was introduced and, consequently, the sensitivity increased from 10 pg to 100 fg. We also reduced a total assay time to 8 h by optimizing hybridization time without compromising test sensitivity. Taken together, our multiplex detection system can improve detection of drug\u2010resistant Gram\u2010positive pathogens from sepsis patients\u2019 blood.<\/jats:p>","DOI":"10.1002\/elps.201600372","type":"journal-article","created":{"date-parts":[[2016,8,30]],"date-time":"2016-08-30T06:55:04Z","timestamp":1472540104000},"page":"3079-3083","source":"Crossref","is-referenced-by-count":4,"title":["Multiplex identification of drug\u2010resistant Gram\u2010positive pathogens using stuffer\u2010free MLPA system"],"prefix":"10.1002","volume":"37","author":[{"given":"Boram","family":"Chung","sequence":"first","affiliation":[{"name":"Department of Microbiology, College of Medicine The Catholic University of Korea Korea  Seoul Korea"},{"name":"Integrated Research Center for Genome Polymorphism, College of Medicine The Catholic University of Korea Korea  Seoul Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chulmin","family":"Park","sequence":"additional","affiliation":[{"name":"Vaccine Bio Research Institute, College of Medicine The Catholic University of Korea Korea  Seoul Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sung\u2010Yeon","family":"Cho","sequence":"additional","affiliation":[{"name":"Vaccine Bio Research Institute, College of Medicine The Catholic University of Korea Korea  Seoul Korea"},{"name":"Division of Infectious Diseases, Department of Internal Medicine, College of Medicine The Catholic University of Korea  Seoul Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sun","family":"Shin","sequence":"additional","affiliation":[{"name":"Department of Microbiology, College of Medicine The Catholic University of Korea Korea  Seoul Korea"},{"name":"Integrated Research Center for Genome Polymorphism, College of Medicine The Catholic University of Korea Korea  Seoul Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seon\u2010Hee","family":"Yim","sequence":"additional","affiliation":[{"name":"Integrated Research Center for Genome Polymorphism, College of Medicine The Catholic University of Korea Korea  Seoul Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gyoo Yeol","family":"Jung","sequence":"additional","affiliation":[{"name":"School of Interdisciplinary Bioscience and Bioengineering Pohang University of Science and Technology  Pohang Gyeongbuk Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dong\u2010Gun","family":"Lee","sequence":"additional","affiliation":[{"name":"Vaccine Bio Research Institute, College of Medicine The Catholic University of Korea Korea  Seoul Korea"},{"name":"Division of Infectious Diseases, Department of Internal Medicine, College of Medicine The Catholic University of Korea  Seoul Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yeun\u2010Jun","family":"Chung","sequence":"additional","affiliation":[{"name":"Department of Microbiology, College of Medicine The Catholic University of Korea Korea  Seoul Korea"},{"name":"Integrated Research Center for Genome Polymorphism, College of Medicine The Catholic University of Korea Korea  Seoul Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2016,9,27]]},"reference":[{"key":"e_1_2_5_2_1","doi-asserted-by":"publisher","DOI":"10.1186\/cc1022"},{"key":"e_1_2_5_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11908-010-0128-x"},{"key":"e_1_2_5_4_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa022139"},{"key":"e_1_2_5_5_1","doi-asserted-by":"publisher","DOI":"10.3947\/ic.2013.45.1.22"},{"key":"e_1_2_5_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mimet.2012.12.012"},{"key":"e_1_2_5_7_1","doi-asserted-by":"publisher","DOI":"10.1556\/EuJMI.3.2013.2.2"},{"key":"e_1_2_5_8_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gnf056"},{"key":"e_1_2_5_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/elps.201200334"},{"key":"e_1_2_5_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2012.02.009"},{"key":"e_1_2_5_11_1","doi-asserted-by":"publisher","DOI":"10.1002\/elps.201300374"},{"key":"e_1_2_5_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/elps.201000091"},{"key":"e_1_2_5_13_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0072349"}],"container-title":["ELECTROPHORESIS"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Felps.201600372","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Felps.201600372","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/elps.201600372","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T14:50:48Z","timestamp":1760539848000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/elps.201600372"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,9,27]]},"references-count":12,"journal-issue":{"issue":"23-24","published-print":{"date-parts":[[2016,12]]}},"alternative-id":["10.1002\/elps.201600372"],"URL":"https:\/\/doi.org\/10.1002\/elps.201600372","archive":["Portico"],"relation":{},"ISSN":["0173-0835","1522-2683"],"issn-type":[{"type":"print","value":"0173-0835"},{"type":"electronic","value":"1522-2683"}],"subject":[],"published":{"date-parts":[[2016,9,27]]}}}