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Development and Evaluation of a Lateral Flow Immunoassay for on-site Diagnosisof Contagious Bovine Pleuropneumonia
KTH, School of Biotechnology (BIO).
2014 (English)Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

The purpose of this study was to develop and evaluate a lateral flow immunoassay, as a point-of-care prototype for diagnosis of contagious bovine pleuropneumonia (CBPP). The assay was based on a panel of recombinant proteins spotted on a nitrocellulose membrane. The study included optimization of assay components, such as running buffer and protein microarray layout. The purpose was to obtain a clear discriminatory capacity between CBPP positive and CBPP negative sera, by developing and evaluating a lateral flow immunoassay that could be used on-site.

The discriminatory capacity between CBPP positive sera and CBPP negative sera in the lateral flow assay was statistically significant with p-values ≤0.05 for recombinant proteins to be printed in microarray spots both individually as well as proteins printed as mixtures. The sensitivity of the assay was 64% and the specificity 100%, which was comparable to current diagnostic methods for CBPP. From these rsults, four combinations of recombinant proteins were selected to print as a microarray, which was based on p-valuesas well as sera coverage. As the purpose of this study was to develop a prototype for on-site usage, a fieldtrip to Nairobi, Kenya, facilitated by the Swedish International Development Cooperation Agency, SIDA, was made to test the prototpyes as well as obtain deeper knowledge of user area. However, the lateral flow immunoassay did not deliver reproducible and stable results during the field trip to Kenya.

Place, publisher, year, edition, pages
Keyword [en]
CBPP, diagnostics, immunoassay, latral flow, recombinant protein
National Category
Engineering and Technology
URN: urn:nbn:se:kth:diva-173620OAI: diva2:854001
Available from: 2015-09-15 Created: 2015-09-15 Last updated: 2015-09-15Bibliographically approved

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