Medizinische Universität Graz - Research portal
Selected Publication:
Föderl-Höbenreich, E.
Development of serodiagnostic tools for melioidosis.
[ Diplomarbeit/Master Thesis (UNI) ] Universität Graz; 2019. pp.59.
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- Authors Med Uni Graz:
- Advisor:
- Wagner-Lichtenegger Gabriel
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- Abstract:
- Melioidosis is a severe infectious disease with a high mortality burden in tropical and subtropical regions and is caused by the gram negative environmental bacterium Burkholderia pseudomallei. Diagnosis is often problematic, due to the rather unspecific symptoms and diagnostic tools with mediocre performance. Up to now, the current diagnostic standard methods are the indirect hemagglutination assay (IHA) and culture, which are both hampered by their low sensitivity (60%). Therefore, there is a great need for the development of new, standardizable and sensitive diagnostic tools. The aim of this work was the development of an improved version of a protein microarray and a point-of-care test for serological testing of melioidosis. The here described protein microarray as well as the lateral flow assay (LFA) are based on a multiplex detection of serum antibodies against B. pseudomallei proteins. On the protein microarray the immune response against up to 400 antigens can be analyzed in parallel, whereas the whole procedure is highly standardizable, which makes it particularly attractive for routine diagnostic. IgG-detection is based on a horse radish peroxidase labeled goat anti-human IgG detection antibody. Based on the data obtained from the improved microarray version and the screening of more than 400 sera, new antigen candidates for the LFA were chosen, to improve its performance. On the LFA, four antigens can be spotted and an individual’s immune response is detected by colloidal gold conjugated secondary anti-human IgG antibodies. The LFA is not only a cost effective, but also a very simple to use point-of-care test, which makes it a perfect tool for an initial diagnosis at the patients home but also for epidemiological studies. In this work it could be demonstrated, that the protein microarray as well as the LFA are easy and fast to perform and provide high sensitivity (89,3%; 70,2%) and specificity (98,1%; 84,7%).