Simple and portable on-site system for nucleic acid-based detection of Clostridium difficile in stool samples using two columns containing microbeads and loop-mediated isothermal amplification
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Baek, Changyoon | - |
dc.contributor.author | Li, Yun Guang | - |
dc.contributor.author | Yoo, Hyun Jin | - |
dc.contributor.author | Cui, Wen Ying | - |
dc.contributor.author | Kang, Jia Tong | - |
dc.contributor.author | Son, Juwon | - |
dc.contributor.author | Min, Junhong | - |
dc.date.accessioned | 2021-11-02T06:40:38Z | - |
dc.date.available | 2021-11-02T06:40:38Z | - |
dc.date.issued | 2022-01 | - |
dc.identifier.issn | 1618-2642 | - |
dc.identifier.issn | 1618-2650 | - |
dc.identifier.uri | https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/50976 | - |
dc.description.abstract | It is challenging to employ nucleic acid-based diagnostics for the in situ detection of Clostridium difficile from complex fecal samples because essential sample preparation and amplification procedures require various experimental resources. In this study, a simple and effective on-site nucleic acid-based detection system was used to detect C. difficile in stool samples. Two columns containing different microbeads, namely, glass and functionalized graphene oxide-coated microbeads, were designed to remove relatively large impurities by filtration and concentrate bacteria, including C. difficile, from stool samples by adsorption. The bacterial nucleic acids were effectively extracted using a small bead beater. The effectiveness of enzyme inhibitors remaining in the sample was efficiently reduced by the direct buffer developed in this study. This sample preparation kit consisting of two simple columns showed better performance in real-time polymerase chain reaction (PCR) and equivalent performance in loop-mediated isothermal amplification (LAMP) than other sample preparation kits, despite 90% simplification of the process. The amplification-ready samples were introduced into two microtubes containing LAMP pre-mixtures (one each for E. coli as an external positive control and C. difficile) by a simple sample loader, which was operated using a syringe. LAMP, which indicates amplification based on color change, was performed at 65 degrees C in a small water bath. The limit of detection (L.O.D) and analytical sensitivity/specificity of our simple and effective kit were compared with those of a commercial kit. C. difficile in stool samples could be detected within 1 h with 10(3) cfu/10 mg using LAMP combined simple on-site detection kit. | - |
dc.format.extent | 9 | - |
dc.language | 영어 | - |
dc.language.iso | ENG | - |
dc.publisher | SPRINGER HEIDELBERG | - |
dc.title | Simple and portable on-site system for nucleic acid-based detection of Clostridium difficile in stool samples using two columns containing microbeads and loop-mediated isothermal amplification | - |
dc.type | Article | - |
dc.identifier.doi | 10.1007/s00216-021-03557-4 | - |
dc.identifier.bibliographicCitation | ANALYTICAL AND BIOANALYTICAL CHEMISTRY, v.414, no.1, pp 613 - 621 | - |
dc.description.isOpenAccess | N | - |
dc.identifier.wosid | 000680299900001 | - |
dc.identifier.scopusid | 2-s2.0-85111806686 | - |
dc.citation.endPage | 621 | - |
dc.citation.number | 1 | - |
dc.citation.startPage | 613 | - |
dc.citation.title | ANALYTICAL AND BIOANALYTICAL CHEMISTRY | - |
dc.citation.volume | 414 | - |
dc.type.docType | Article | - |
dc.publisher.location | 독일 | - |
dc.subject.keywordAuthor | Clostridium difficile | - |
dc.subject.keywordAuthor | Stool sample | - |
dc.subject.keywordAuthor | Graphene oxide | - |
dc.subject.keywordAuthor | Portable on-site system | - |
dc.subject.keywordAuthor | Isothermal amplification | - |
dc.subject.keywordPlus | TOXIN-B | - |
dc.subject.keywordPlus | RAPID DETECTION | - |
dc.relation.journalResearchArea | Biochemistry & Molecular Biology | - |
dc.relation.journalResearchArea | Chemistry | - |
dc.relation.journalWebOfScienceCategory | Biochemical Research Methods | - |
dc.relation.journalWebOfScienceCategory | Chemistry, Analytical | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
Items in ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.
84, Heukseok-ro, Dongjak-gu, Seoul, Republic of Korea (06974)02-820-6194
COPYRIGHT 2019 Chung-Ang University All Rights Reserved.
Certain data included herein are derived from the © Web of Science of Clarivate Analytics. All rights reserved.
You may not copy or re-distribute this material in whole or in part without the prior written consent of Clarivate Analytics.