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Rapid signal enhancement method for nanoprobe-based biosensing
KTH, Skolan för bioteknologi (BIO), Proteomik och nanobioteknologi.
KTH, Skolan för bioteknologi (BIO), Proteomik och nanobioteknologi.
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2017 (Engelska)Ingår i: Scientific Reports, E-ISSN 2045-2322, Vol. 7, artikel-id 6837Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

The introduction of nanomaterials as detection reagents has enabled improved sensitivity and facilitated detection in a variety of bioanalytical assays. However, high nanoprobe densities are typically needed for colorimetric detection and to circumvent this limitation several enhancement protocols have been reported. Nevertheless, there is currently a lack of universal, enzyme-free and versatile methods that can be readily applied to existing as well as new biosensing strategies. The novel method presented here is shown to enhance the signal of gold nanoparticles enabling visual detection of a spot containing < 10 nanoparticles. Detection of Protein G on paper arrays was improved by a 100-fold amplification factor in under five minutes of assay time, using IgG-labelled gold, silver, silica and iron oxide nanoprobes. Furthermore, we show that the presented protocol can be applied to a commercial allergen microarray assay, ImmunoCAP ISAC sIgE 112, attaining a good agreement with fluorescent detection when analysing human clinical samples.

Ort, förlag, år, upplaga, sidor
NATURE PUBLISHING GROUP , 2017. Vol. 7, artikel-id 6837
Nationell ämneskategori
Analytisk kemi
Identifikatorer
URN: urn:nbn:se:kth:diva-212341DOI: 10.1038/s41598-017-07030-0ISI: 000406610000088PubMedID: 28754966Scopus ID: 2-s2.0-85026428495OAI: oai:DiVA.org:kth-212341DiVA, id: diva2:1135494
Anmärkning

QC 20170823

Tillgänglig från: 2017-08-23 Skapad: 2017-08-23 Senast uppdaterad: 2024-03-18Bibliografiskt granskad
Ingår i avhandling
1. Novel planar and particle-based microarrays for point-of-care diagnostics
Öppna denna publikation i ny flik eller fönster >>Novel planar and particle-based microarrays for point-of-care diagnostics
2019 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Abstract [en]

Point-of-care assays are easy-to-use, portable and inexpensive tests that can

be used to aid diagnostics by measuring levels of disease-specific molecules

in settings where access to advanced laboratory equipment and trained

personnel are limited, such as at the patient's bedside or in low resource

parts of developing countries. In order to achieve high multiplexing

capacities, such assays can be based on planar microarrays consisting of

spots immobilized on a flat surface or on particle-based microarrays based

on populations of encoded particles. The aim of the work presented in this

thesis is to develop new point-of-care amenable planar and particle-based

microarrays that allow for highly multiplexed assays while maintaining low

sample-to-result times, complexity and instrumentation requirements.

Paper I demonstrates the use graphically encoded particles for colorimetric

detection of autoantibodies using a consumer-grade flatbed scanner. Four

graphical characters on the surface of each particle allows for millions of

codes and the use of gold nanoparticles as a detection label allows both the

code and the signal intensity to be read out in a single image.

Paper II describes a signal enhancement method that increases the

sensitivity of gold nanoparticle detection on planar microarrays. Using this

method, detection of allergen-specific IgE can be carried out using a

consumer-grade flatbed scanner instead of a more expensive fluorescence

scanner without sacrificing assay performance.

Paper III demonstrates the use of an isothermal DNA amplification method

for detection of adenoviral DNA on a paper-based microarray. Using an

isothermal amplification method eliminates the need for a thermocycler,

reducing the instrumentation required for such detection.

Paper IV shows the use of solid-phase PCR to amplify bacterial DNA directly

on the surface of particles. This strategy reduces assay time by eliminating

the need for separate amplification and hybridisation steps.

Ort, förlag, år, upplaga, sidor
Stockholm: KTH Royal Institute of Technology, 2019. s. 88
Serie
TRITA-CBH-FOU ; 2019:13
Nyckelord
Planar microarrays, Particle-based microarrays, Point-of-care diagnostics, Colorimetric detection, Signal enhancement, Isothermal DNA amplification, Solid-phase DNA amplification
Nationell ämneskategori
Läkemedel- och medicinsk processbioteknik
Forskningsämne
Bioteknologi
Identifikatorer
urn:nbn:se:kth:diva-244430 (URN)978-91-7873-101-5 (ISBN)
Disputation
2019-03-22, Science for Life Laboratory, room Air & Fire, Tomtebodavägen 23A, Solna, 10:00 (Engelska)
Opponent
Handledare
Anmärkning

QC 20190221

Tillgänglig från: 2019-02-21 Skapad: 2019-02-20 Senast uppdaterad: 2025-02-17Bibliografiskt granskad

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Dias, Jorge TSvedberg, GustavGantelius, Jesper

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