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A lateral flow protein microarray for rapid determination of contagious bovine pleuropneumonia status in bovine serum
KTH, School of Biotechnology (BIO), Nano Biotechnology.
KTH, School of Biotechnology (BIO), Proteomics.ORCID iD: 0000-0001-8141-8449
KTH, School of Biotechnology (BIO).
KTH, School of Biotechnology (BIO), Proteomics.ORCID iD: 0000-0002-1855-703X
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2010 (English)In: Journal of Microbiological Methods, ISSN 0167-7012, E-ISSN 1872-8359, Vol. 82, no 1, 11-18 p.Article in journal (Refereed) Published
Abstract [en]

Novel analytical methods for a next generation of diagnostic devices combine attributes from sensitive, accurate, fast, simple and multiplexed analysis methods. Here, we describe a possible contribution to these by the application of a lateral flow microarray where a panel of recombinant protein antigens was used to differentiate bovine serum samples in the context of the lung disease contagious bovine pleuropneumonia (CBPP). Lateral flow arrays were produced by attaching nitrocellulose onto microscopic slides and spotting of the recombinant proteins onto the membranes. The developed assay included evaluations of substrate matrix and detection reagents to allow for short assay times and convenient read-out options, and to yield a total assay time from sample application to data acquisition of less than ten minutes. It was found that healthy and disease-affected animals could be discriminated (AUC = 97%), and we suggest that the use of an antigen panel in combination with the lateral flow device offers an emerging analytical tool towards a simplified but accurate on-site diagnosis.

Place, publisher, year, edition, pages
2010. Vol. 82, no 1, 11-18 p.
Keyword [en]
Lateral flow, Microarray, Gold nanobeads, Contagious bovine pleuropneumonia, Serum analysis, Multiplex, Antigens
National Category
Industrial Biotechnology
URN: urn:nbn:se:kth:diva-14099DOI: 10.1016/j.mimet.2010.03.007ISI: 000279567700002ScopusID: 2-s2.0-77953541930OAI: diva2:329732
QC 20100713Available from: 2010-07-13 Created: 2010-07-13 Last updated: 2011-02-02Bibliographically approved
In thesis
1. Novel diagnostic microarray assay formats towards comprehensive on-site analysis
Open this publication in new window or tab >>Novel diagnostic microarray assay formats towards comprehensive on-site analysis
2009 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Advances in molecular methods for analyzing DNA, RNA and proteins in humans as well as in other animals, plants, fungi, bacteria or viruses have greatly increased the resolution with which we can study life’s complexity and dynamics on earth. While genomic, transcriptomic and proteomic laboratory tools for molecular diagnosis of disease are rapidly becoming more comprehensive, the access to such advanced yet often expensive and centralized procedures is limited. There is a great need for rapid and comprehensive diagnostic methods in low-resource settings or contexts where a person can not or will not go to a hospital or medical laboratory, yet where a clinical analysis is urgent.

In this thesis, results from development and characterization of novel technologies for DNA and protein microarray analysis are presented. Emphasis is on methods that could provide rapid, cost-effective and portable analysis with convenient readout and retained diagnostic accuracy. The first study presents a magnetic bead-based approach for DNA microarray analysis for a rapid visual detection of single nucleotide polymorphisms. In the second work, magnetic beads were used as detection reagents for rapid differential detection of presence of pestiviral family members using a DNA oligonucleotide microarray with read-out by means of a tabletop scanner or a digital camera. In paper three, autoimmune responses from human sera were detected on a protein autoantigen microarray, again by means of magnetic bead analysis. Here, special emphasis was made in comprehensively comparing the performance of the magnetic bead detection to common fluorescence-based detection. In the fourth study, an immunochromatographic lateral flow protein microarray assay is presented for application in the classification of contagious pleuropneumonia from bovine serum samples. The analysis could be performed within 10 minutes using a table top scanner, and the performance of the assay was shown to be comparable to that of a cocktail ELISA. In the fifth paper, the lateral flow microarray framework is investigated in further detail by means of experiments and numerical simulation. It was found that downstream effects play an important role, and the results further suggest that the downstream binding profiles may find use in simple affinity evaluation.

Place, publisher, year, edition, pages
Stockholm: KTH, 2009. xii, 90 p.
Trita-BIO-Report, ISSN 1654-2312 ; 2009:18
magnetic microbeads, gold nanobeads, microarrays, lateral flow, on-site diagnosis, SNPs, pestivirus, autoimmunity, contagious bovine pleuropneumonia, comsol multiphysics, finite element method
National Category
Microbiology Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
urn:nbn:se:kth:diva-11221 (URN)978-91-7415-416-0 (ISBN)
Public defence
2009-10-23, FR4 Oskar Klein, AlbaNova Universitetscentrum, Roslagstullsbacken, Stockholm, 10:00 (English)
QC 20100713Available from: 2009-10-07 Created: 2009-10-07 Last updated: 2011-11-23Bibliographically approved

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