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Developing liquid AP-MALDI MS as a rapid diagnostics method for bovine tuberculosis - SE3327

In this project a novel diagnostic method will be developed for the rapid, large-scale detection of bovine tuberculosis (bTB) using mass spectrometry (MS) profiling of liquid biopsies (animal biofluids). We will exploit the knowledge obtained from a milk analysis approach that has recently been highly successful for the (early) detection of bovine mastitis. The aim is to develop a test using just a few microlitres of biofluid that can be easily and non-invasively collected and rapidly analysed in test centres, which can provide an analysis turn-around time of minutes once the samples are received. Future testing platforms could also be set up on larger farms as the methodology is capable of multiplexing (able to detect a multitude of diseases), making it universal and highly cost-effective, justifying the capital cost for farm-side analysis.
Profiling of the biomolecular content of liquid biopsies can be highly specific and sensitive in the detection of disease. Modern MS has become the method of choice in detecting thousands of different biomolecules, and thus potential disease markers, from liquid biopsies, such as blood, milk or saliva, in a single analysis. These analyses typically require extensive sample preparation and extended analysis time for each individual biopsy specimen. We have recently developed a new MS method, called liquid atmospheric pressure (AP) matrix-assisted laser desorption/ionisation (MALDI) MS, for profiling milk from dairy cows that can detect mastitis with extremely high accuracy (>99%). This method has even been able to detect bovine mastitis 2 days before its detection by current conventional testing with good accuracy (99% specificity, 50% sensitivity). Important aspects of the new methodology are its speed and scalability, allowing thousands of samples to be measured per day on a single platform, and its extremely simple sample preparation, requiring only two quick steps that can be performed within the same sample preparation vial.
The proposed research will utilise easy collection of liquid biopsies, mainly saliva or nasal fluids, and exploit the latest in liquid AP-MALDI MS profiling. The former will make specimen collection a fast process that can be implemented on any farm. The latter will provide an analytical tool that can be extremely fast and large-scale in its analysis while being highly accurate. We will focus on the biomolecular classes that have previously been shown to be sufficiently abundant in the respective biofluid, starting with lipids in saliva or nasal fluids. Samples will be obtained from the research herd of CEDAR (Centre of Dairy Research) at the University of Reading and other local herds. Sample collection will include specimens that will allow bTB to be distinguished from other infections at the acute phase.

The aim of the proposed project is to develop a mass spectrometry (MS)-based rapid diagnostics tool for bovine tubercolusis (bTB) employing a liquid biopsy specimen collection that is user-friendly and non-invasive. The important and unique features of this tool will be (i) its speed allowing bTB detection – from specimen collection to bTB detection – within minutes (or a few hours if sent to a laboratory) and (ii) its simplicity and cost-effectiveness using inexpensive consumables and an easy protocol for farm-side, non-invasive specimen collection. Both aspects will allow large-scale and frequent specimen analyses, thus allowing continuous (at a much higher frequency than currently employed) and population-wide screening with the potential to achieve Defra TB programme’s objective to eradicate bTB in England by 2038. The development of this tool includes innovative research, employing novel liquid AP-MALDI MS profiling analysis, leading to a product/method prototype that can support bTB stakeholders in rapidly diagnosing bTB infections.

Research objectives will include (A) identifying the body fluid that can provide the greatest number of liquid AP-MALDI MS-detectable diagnostic bTB-specific biomarkers and developing its simple, non-invasive and user-friendly collection, (B) optimising the extraction of diagnostically important bTB biomarkers from the liquid biopsy and their liquid AP-MALDI MS sample preparation and (C) optimising the MS recording parameters and the subsequent data mining for best bTB detection model building.

Time-Scale and Cost
From: 2021

To: 2022

Cost: £99,613
Contractor / Funded Organisations
University of Reading