DNA & Protein detection based on microbead agglutination

Handle URI:
http://hdl.handle.net/10754/303788
Title:
DNA & Protein detection based on microbead agglutination
Authors:
Kodzius, Rimantas ( 0000-0001-9417-8894 ) ; Castro, David; Foulds, Ian G.; Parameswaran, Ash M.; Sumanpreet, K. Chhina
Abstract:
We report a simple and rapid room temperature assay for point-of-care (POC) testing that is based on specific agglutination. Agglutination tests are based on aggregation of microparticles in the presence of a specific analyte thus enabling the macroscopic observation. Agglutination-based tests are most often used to explore the antibody-antigen reactions. Agglutination has been used for mode protein assays using a biotin/streptavidin two-component system, as well as a hybridization based two-component assay; however, as our work shows, two-component systems are prone to self-termination of the linking analyte and thus have a lower sensitivity. Three component systems have also been used with DNA hybridization, as in our work; however, their assay requires 48 hours for incubation, while our assay is performed in 5 minutes making it a real candidate for POC testing. We demonstrate three assays: a two-component biotin/streptavidin assay, a three-component hybridization assay using single stranded DNA (ssDNA) molecules and a stepped three-component hybridization assay. The comparison of these three assays shows our simple stepped three-component agglutination assay to be rapid at room temperature and more sensitive than the two-component version by an order of magnitude. An agglutination assay was also performed in a PDMS microfluidic chip where agglutinated beads were trapped by filter columns for easy observation. We developed a rapid (5 minute) room temperature assay, which is based on microbead agglutination. Our three-component assay solves the linker self-termination issue allowing an order of magnitude increase in sensitivity over two–component assays. Our stepped version of the three-component assay solves the issue with probe site saturation thus enabling a wider range of detection. Detection of the agglutinated beads with the naked eye by trapping in microfluidic channels has been shown.
KAUST Department:
Computational Bioscience Research Center (CBRC)
Publisher:
Hilton Head Workshop on the science and technology of solid-state sensors, actuators, and microsystems
Journal:
Hilton Head Workshop on the science and technology of solid-state sensors, actuators, and microsystems
Issue Date:
6-Jun-2012
Type:
Poster
Additional Links:
http://www.transducer-research-foundation.org/archive/hh2012/program/HH2012_FinalProgram.pdf
Appears in Collections:
Posters; Computational Bioscience Research Center (CBRC)

Full metadata record

DC FieldValue Language
dc.contributor.authorKodzius, Rimantasen
dc.contributor.authorCastro, Daviden
dc.contributor.authorFoulds, Ian G.en
dc.contributor.authorParameswaran, Ash M.en
dc.contributor.authorSumanpreet, K. Chhinaen
dc.date.accessioned2013-10-22T08:41:52Z-
dc.date.available2013-10-22T08:41:52Z-
dc.date.issued2012-06-06en
dc.identifier.urihttp://hdl.handle.net/10754/303788en
dc.description.abstractWe report a simple and rapid room temperature assay for point-of-care (POC) testing that is based on specific agglutination. Agglutination tests are based on aggregation of microparticles in the presence of a specific analyte thus enabling the macroscopic observation. Agglutination-based tests are most often used to explore the antibody-antigen reactions. Agglutination has been used for mode protein assays using a biotin/streptavidin two-component system, as well as a hybridization based two-component assay; however, as our work shows, two-component systems are prone to self-termination of the linking analyte and thus have a lower sensitivity. Three component systems have also been used with DNA hybridization, as in our work; however, their assay requires 48 hours for incubation, while our assay is performed in 5 minutes making it a real candidate for POC testing. We demonstrate three assays: a two-component biotin/streptavidin assay, a three-component hybridization assay using single stranded DNA (ssDNA) molecules and a stepped three-component hybridization assay. The comparison of these three assays shows our simple stepped three-component agglutination assay to be rapid at room temperature and more sensitive than the two-component version by an order of magnitude. An agglutination assay was also performed in a PDMS microfluidic chip where agglutinated beads were trapped by filter columns for easy observation. We developed a rapid (5 minute) room temperature assay, which is based on microbead agglutination. Our three-component assay solves the linker self-termination issue allowing an order of magnitude increase in sensitivity over two–component assays. Our stepped version of the three-component assay solves the issue with probe site saturation thus enabling a wider range of detection. Detection of the agglutinated beads with the naked eye by trapping in microfluidic channels has been shown.en
dc.language.isoenen
dc.publisherHilton Head Workshop on the science and technology of solid-state sensors, actuators, and microsystemsen
dc.relation.urlhttp://www.transducer-research-foundation.org/archive/hh2012/program/HH2012_FinalProgram.pdfen
dc.subjectMicrofluidicsen
dc.subjectAgglutinationen
dc.subjectMicrobeadsen
dc.subjectDNA hybridizationen
dc.subjectBiotinylated BSAen
dc.titleDNA & Protein detection based on microbead agglutinationen
dc.typePosteren
dc.contributor.departmentComputational Bioscience Research Center (CBRC)en
dc.identifier.journalHilton Head Workshop on the science and technology of solid-state sensors, actuators, and microsystemsen
dc.eprint.versionPublisher's Version/PDFen
dc.contributor.affiliationKing Abdullah University of Science and Technology (KAUST)en
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