Rapid visual identification of PCR amplified nucleic acids by centrifugal gel separation: Potential use for molecular point-of-care tests
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Hwang, Sang-Hyun | - |
dc.contributor.author | Kim, Dong-Eun | - |
dc.contributor.author | Im, Ji-Hyun | - |
dc.contributor.author | Kang, Su-Jin | - |
dc.contributor.author | Lee, Do-Hoon | - |
dc.contributor.author | Son, Sang Jun | - |
dc.date.available | 2020-02-28T01:45:49Z | - |
dc.date.created | 2020-02-06 | - |
dc.date.issued | 2016-05-15 | - |
dc.identifier.issn | 0956-5663 | - |
dc.identifier.uri | https://scholarworks.bwise.kr/gachon/handle/2020.sw.gachon/8294 | - |
dc.description.abstract | Recently, nucleic acid amplification and detection techniques have progressed based on advances in in microfluidics, microelectronics, and optical systems. Nucleic acids amplification based point-of-care test (POCT) in resource-limited settings requires simple visual detection methods. Several biosensing methods including lateral flow immunoassays (LFIA) were previously used to visually detect nucleic acids. However, prolonged assay time, several washing steps, and a need for specific antibodies limited their use. Here we developed a novel, rapid method to visualize amplified nucleic acids with naked eyes in clinical samples. First, we optimized conditions based on separation using very low centrifugal force and a density medium to detect human papillomavirus (HPV)-16 DNA in cervical specimens. After DNA extraction, HPV16 PCR was performed with biotin-labeled forward primer and Cy3-labeled reverse primer. PCR amplicon was mixed with streptavidin-magnetic beads, introduced into the density medium. After two-minute centrifugation, the result was visually identified. This system showed identical results with commercial HPV real-time PCR for 30 clinical samples and could detect up to 10(2) copies/mL of HPV DNA without any optical instruments. This robust and sensitive visual detection system is suitable for non-specialist personnel and point-of-care diagnosis in low-resource settings. (C) 2016 Elsevier B.V. All rights reserved. | - |
dc.language | 영어 | - |
dc.language.iso | en | - |
dc.publisher | ELSEVIER ADVANCED TECHNOLOGY | - |
dc.relation.isPartOf | BIOSENSORS & BIOELECTRONICS | - |
dc.subject | FLUORESCENT DYES | - |
dc.subject | DNA | - |
dc.title | Rapid visual identification of PCR amplified nucleic acids by centrifugal gel separation: Potential use for molecular point-of-care tests | - |
dc.type | Article | - |
dc.type.rims | ART | - |
dc.description.journalClass | 1 | - |
dc.identifier.wosid | 000370309200113 | - |
dc.identifier.doi | 10.1016/j.bios.2016.01.006 | - |
dc.identifier.bibliographicCitation | BIOSENSORS & BIOELECTRONICS, v.79, pp.829 - 834 | - |
dc.identifier.scopusid | 2-s2.0-84954271773 | - |
dc.citation.endPage | 834 | - |
dc.citation.startPage | 829 | - |
dc.citation.title | BIOSENSORS & BIOELECTRONICS | - |
dc.citation.volume | 79 | - |
dc.contributor.affiliatedAuthor | Son, Sang Jun | - |
dc.type.docType | Article | - |
dc.subject.keywordAuthor | DNA separation | - |
dc.subject.keywordAuthor | Molecular POC | - |
dc.subject.keywordAuthor | Biosensing | - |
dc.subject.keywordAuthor | Gel-separation | - |
dc.subject.keywordAuthor | Centrifugation | - |
dc.subject.keywordAuthor | Visual detection | - |
dc.subject.keywordPlus | FLUORESCENT DYES | - |
dc.subject.keywordPlus | DNA | - |
dc.relation.journalResearchArea | Biophysics | - |
dc.relation.journalResearchArea | Biotechnology & Applied Microbiology | - |
dc.relation.journalResearchArea | Chemistry | - |
dc.relation.journalResearchArea | Electrochemistry | - |
dc.relation.journalResearchArea | Science & Technology - Other Topics | - |
dc.relation.journalWebOfScienceCategory | Biophysics | - |
dc.relation.journalWebOfScienceCategory | Biotechnology & Applied Microbiology | - |
dc.relation.journalWebOfScienceCategory | Chemistry, Analytical | - |
dc.relation.journalWebOfScienceCategory | Electrochemistry | - |
dc.relation.journalWebOfScienceCategory | Nanoscience & Nanotechnology | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
Items in ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.
1342, Seongnam-daero, Sujeong-gu, Seongnam-si, Gyeonggi-do, Republic of Korea(13120)031-750-5114
COPYRIGHT 2020 Gachon 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.