Proximity Ligation Assay for the specific detection of Aspergillus species

Gemma Johnson1, Mark Shannon2, Christopher Thornton3, Samir Agrawal4, Cornelia Lass-Flörl5, Wolfgang Mutschlechner5, Stephen Bustin1
1Postgraduate Medical Institute, Faculty of Medical Science, Anglia Ruskin University, UK; 
2Thermo Fisher Scientific, USA; 
3School of Biosciences, University of Exeter, UK; 
4Blizard Institute of Cellular and Molecular Science, Queen Mary University, London, UK;
5Division of Hygiene and Medical Microbiology, Innsbruck Medical University, Innsbruck, Austria

Antibody- and real-time quantitative (qPCR)-based assays are proving useful for a more sensitive and specific identification ofAspergillus in clinical samples. However, both approaches have important shortcomings: the relative insensitivity of current antibody-based strategies makes them less suitable for earliest possible diagnosis, whereas the mere detection of pathogen DNA is uninformative with regards to its viability or infectivity. Hence the ultimate assay would combine the specificity of pathogen-specific antibody detection with the sensitivity of qPCR. The proximity ligation assay (PLA) uses qPCR to detect the interaction of antibodies with their specific antigens.
We have used this technology with an Aspergillus specific monoclonal antibody (MAb) to demonstrate sensitive and specific detection ofAspergillus mannoprotein in clinically relevant samples. The assay is simple to use, rapid and significantly more sensitive than other antibody-based assays in current use. This has important implications for early diagnosis and targeted treatment of Invasive Aspergillosis.

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