DC Field | Value | Language |
---|---|---|
dc.contributor.author | Chung, B | - |
dc.contributor.author | Park, C | - |
dc.contributor.author | Cho, SY | - |
dc.contributor.author | Shin, S | - |
dc.contributor.author | Yim, SH | - |
dc.contributor.author | Jung, GY | - |
dc.contributor.author | Lee, DG | - |
dc.contributor.author | Chung, YJ | - |
dc.date.accessioned | 2018-01-04T10:57:04Z | - |
dc.date.available | 2018-01-04T10:57:04Z | - |
dc.date.created | 2017-07-05 | - |
dc.date.issued | 2016-12 | - |
dc.identifier.issn | 0173-0835 | - |
dc.identifier.uri | https://oasis.postech.ac.kr/handle/2014.oak/39225 | - |
dc.description.abstract | Early detection of pathogens from blood and identification of their drug resistance are essential for sepsis management. However, conventional culture-based methods require relatively longer time to identify drug-resistant pathogens, which delays therapeutic decisions. For precise multiplex detection of drug-resistant Gram-positive pathogens, we developed a method by using stuffer-free multiplex ligation-dependent probe amplification (MLPA) coupled with high-resolution CE single-strand conformation polymorphisms (CE-SSCP) system. We designed three probe sets for genes specific to Gram-positive species (Staphylococcus aureus: nuc, Enterococcus faecium: sodA, and Streptococcus pneumoniae: lytA) and two sets for genes associated with drug resistance (mecA and vanA) to discriminate major Gram-positive 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-specific peaks were successfully observed for all the strains. To improve sensitivity of the method, a target-specific 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-resistant Gram-positive pathogens from sepsis patients' blood. | - |
dc.language | English | - |
dc.publisher | WILEY-BLACKWELL | - |
dc.relation.isPartOf | ELECTROPHORESIS | - |
dc.title | Multiplex identification of drug-resistant Gram-positive pathogens using stuffer-free MLPA system | - |
dc.type | Article | - |
dc.identifier.doi | 10.1002/ELPS.201600372 | - |
dc.type.rims | ART | - |
dc.identifier.bibliographicCitation | ELECTROPHORESIS, v.37, no.23-24, pp.3079 - 3083 | - |
dc.identifier.wosid | 000392421400006 | - |
dc.date.tcdate | 2019-02-01 | - |
dc.citation.endPage | 3083 | - |
dc.citation.number | 23-24 | - |
dc.citation.startPage | 3079 | - |
dc.citation.title | ELECTROPHORESIS | - |
dc.citation.volume | 37 | - |
dc.contributor.affiliatedAuthor | Jung, GY | - |
dc.identifier.scopusid | 2-s2.0-84992450230 | - |
dc.description.journalClass | 1 | - |
dc.description.journalClass | 1 | - |
dc.description.wostc | 1 | - |
dc.description.scptc | 1 | * |
dc.date.scptcdate | 2018-05-121 | * |
dc.description.isOpenAccess | N | - |
dc.type.docType | Article | - |
dc.subject.keywordAuthor | Capillary electrophoresis | - |
dc.subject.keywordAuthor | Gram-positive pathogen | - |
dc.subject.keywordAuthor | Multiplex ligation-dependent probe amplification | - |
dc.subject.keywordAuthor | Sepsis | - |
dc.relation.journalWebOfScienceCategory | Biochemical Research Methods | - |
dc.relation.journalWebOfScienceCategory | Chemistry, Analytical | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
dc.relation.journalResearchArea | Biochemistry & Molecular Biology | - |
dc.relation.journalResearchArea | Chemistry | - |
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