Title Promoter Affiliations Abstract "Mast cell activation tests (MAT): from an innovative diagnostic to the expansion of a service facility." "Didier Ebo" Immunology "Drug hypersensitivity constitutes a significant health problem with serious consequences of both mis- and overdiagnosis. nfortunately, correct diagnosis of drug hypersensitivity can pose significant challenges, mainly because of our knowledge gaps in the molecular and pathophysiological processes underlying immediate and non-immediate drug hypersensitivity reactions but also, most importantly, because of the unavailability of reliable in vitro tests and uncertainties associated with skin testing 1-3. This ""proof-of-concept project"" is an apex of our ongoing overarching research on the cellular processes, pathophysiology and diagnosis of immediate drug hypersensitivity reactions resulting from mast cell and basophil degranulation 1-30. Likely, this project will deepen our insights and shift paradigms in the mechanisms that govern mast cell degranulation finally culminating in immediate drug hypersensitivity reactions. However, our proposal primarily focuses on i) the confirmation of our innovative MAT (mast cell activation test) to be a performant diagnostic and ii) to expand capacity of our service platform to offer these tests to colleagues and industry. With respect to the valorisation, this POC-proposal should promote our services and offering our MAT (amongst other diagnostics) to more colleagues and industry. Today, our laboratory has already created a restricted service platform. In other words, this POC-project will not restrict to confirmation of the MAT, but will also invest, via several initiatives, in the expansion and promotion of our services (inter)nationally, application for a full patent, and in the set-up of a business model. In this context, communication of the data, analyses of demands and market studies are critical. Clearly, the potential of the MAT, which we already filed a priority application for and necessitates only patients' sera, is broad. Moreover, preliminary results from our market analysis, and the potential of additional indications, could result in industrial interest and the creation of a spin-off." "Mass spectrometry-based autoantibody tests for specific diagnosis of autoimmune diseases" "Xavier Bossuyt" "Clinical and Diagnostic Immunology, Laboratory for Clinical Infectious and Inflammatory Disorders, Immunobiology (Rega Institute), Laboratory of Protein Phosphorylation and Proteomics, Skeletal Biology and Engineering Research Center, Laboratory for Muscle Diseases and Neuropathies" "Autoimmune diseases are chronic conditions that collectively affect approximately 1 in 10 people in Europe and the US. They are considered as one of the leading causes of mortality in young and middle-aged women. Anti-Nuclear Antibody (ANA) testing is helpful for the clinical diagnosis and classification of a number of autoimmune diseases including systemic lupus erythematosus (SLE), Sjögren’s syndrome (SS), systemic sclerosis (SSc), mixed connective tissue disease (MCTD) and idiopathic inflammatory myopathies (IIM). The global ANA testing market is experiencing a rapid growth due to rising prevalence of autoimmune disorders and increasing demand for more advanced test solutions. Indeed, individuals who develop an organ-specific autoimmune disease have a high likelihood of developing additional autoimmune disorders. There is a need for new technologies that can evaluate multiple immune reactivities in a small sample volume. One of the major factors hindering the growth of this market are the limitations of the current autoimmune disease diagnostics in terms of accuracy and specificity. Successful tests are those that allow rapid decision making and administration of suitable therapies for treating individuals with autoimmune diseases, especially with the approval of more and more targeted therapies. The identification of novel biomarkers and setup of sensitive tests for simultaneous measurement of multiple autoantibodies are key to reach these goals. KU Leuven is at the forefront of clinical diagnosis of autoimmune diseases and has recently demonstrated proof-of-concept for the use of an in-house developed mass-spectrometry based method for identification of autoantibodies in rheumatic diseases. Such mass spectrometry-based test has the great advantage that it is highly specific, that is allows easy multiplexing and that, in contrast to all other tests available, it measures the interaction between antigen and antibody in fluid phase, which is of utmost importance." "A way out of the reagents shortages: an MS-based diagnostic test focused on corona protein detection" "Dieter Deforce" "Department of Biomolecular Medicine, Department of Pharmaceutics" "CoVid-19 patient diagnosis is currently performed almost exclusively by qPCR-based approaches. This is leading to a world wide restraint on patient testing and a global shortage in reagents required for such assays. We here propose to detect corona proteins instead, using mass spectrometry. We have already proved its applicability, and now need to scale up the assay." "Improving the guidance and support of transfer students in engineering: A validated diagnostic test and effective interventions" "Greet Langie" "Mechanical Engineering Technology, De Nayer (Sint-Katelijne-Waver) Campus, School Psychology and Development in Context, Production Engineering, Machine Design and Automation (PMA) Section" "In order to stimulate flexible lifelong learning, the educational system of Flanders provides alternative ways to enter a Master’s programme, next to the traditional academic Bachelor’s programme. Students who obtained a professional Bachelor’s degree can enrol in an academic Master’s programme provided that they successfully complete a transfer programme. In general, these  transfer students enter university for the first time, just like traditional first-year students. Transfer students at the Faculty of Engineering Technology (FET) experience similar adaptation problems as the first-year students at FET. However, transfer students feel significantly less prepared. Nevertheless, their outcome variables (i.e. academic achievement and dropout rate) after one year of enrolment are similar. Approximately 50% of the transfer students drop out. This high dropout rate is the principal rationale for this research. It is of paramount importance to improve the guidance of students in their educational choice before enrolment as well as to provide them with the required support once they are enrolled. In order to achieve this, the following steps are performed: (1) development of a validated diagnostic test and (2) development and implementation of effective interventions (i.e. a student support programme).The diagnostic test is voluntary, non-binding (i.e. if students do not pass the test, they can still enrol in the programme), and preferably organised before enrolment in the transfer programme. The objectives of the test are (1) to provide students with feedback about their skills and capacities thus to stimulate them to make a well-considered educational choice; and (2) to encourage students to participate in interventions before or during their transfer programme in order to overcome stumbling blocks. The development of the diagnostic test was an iterative process over multiple years, of which the first version was based on (1) an analysis of a diagnostic test for the regular first-year students and (2) students’ stumbling blocks, mathematics and study strategies, which were defined during focus group discussions. It is important that the test includes both cognitive and non-cognitive tests, since students need to realise that both are important for study success at university.Given that the aim is to properly inform students before enrolment and support them after enrolment, two types of advisory models, based on pre-entry characteristics, are distinguished. The first model, a students’ background model, only includes fixed variables (i.e. prior schooling). This model explains a substantial part of variation in students’ grades, but since these variables are fixed, the students’ background model is primarily useful before enrolment. The second model, a diagnostic model, only includes malleable variables (i.e. skills and abilities) that are measured in the diagnostic test. Although this model does not explain the same amount of variance in students’ grades as the students’ background model, the diagnostic model is useful both before and after enrolment. When students receive actionable feedback about their test results and are given the opportunity to participate in interventions, they can enhance their skills and abilities, which in turn can improve students’ academic achievement.Eight interventions were developed in this dissertation and combined into a student support programme. This student support programme starts in the third year of the professional Bachelor’s programme and ends after the transfer programme. The student support programme aims to (1) attract the right students, (2) decrease the feeling of unpreparedness at the beginning of the academic year, and (3) support students after enrolment. Both the effectiveness and efficiency of the eight interventions are examined. The results are combined into an effectiveness/efficiency matrix. This study shows that the most effective interventions are not always the most efficient and vice versa. For two interventions that focus on the students’ stumbling blocks, mathematics and study strategies, a more in-depth analysis of the effectiveness is performed. In this study there was significant evidence for the effectiveness of the mathematics MOOC." "Towards improved monitoring of human soil-transmitted helminthiases by serology: Translating a proven veterinary diagnostic test to human medicine" "Peter Geldhof" "Department of Translational Physiology, Infectiology and Public Health, Jimma University" "In pigs, we showed that serology has a clear added value in the detection of soil-transmitted helminths and estimating the impact of infections in these animals. In this project, we want to evaluate whether our serological test also provide additional insights into the epidemiology of soil-transmitted helminth infections in humans and whether it can be used to monitor deworming programs." "Development of a minimally-invasive test for accurate diagnosis of Cancer of Unknown Primary (CUP) using DNA methylation profiling" "Katleen De Preter" "Department of Biochemistry and microbiology, Department of Internal Medicine and Pediatrics, Department of Diagnostic Sciences, Department of Biomolecular Medicine" "Three to five percent of people who are confronted with cancer will be diagnosed with Cancer of Unknown Primary (CUP); a metastasized cancer of which the tissue-of-origin cannot be determined. If possible, a biopsy of the tumor will be taken for pathological examination, but this can take up to two weeks to plan, execute and analyze. Diagnosis usually requires multiple rounds of immunostaining, a slow and costly method that leaves about 34% of CUPs without definite diagnosis. Because treatment options are limited to aspecific chemotherapy, CUP is the fourth most common cause of cancer death.The tumor DNA methylation pattern is a unique ‘fingerprint’ that can be used to determine the tissue-of-origin in CUPs. A novel technique, called cfRRBS, developed at the UGent-VIB allows the identification of the methylation profile starting from minimal amounts of highly fragmented DNA. With this technique not only DNA extracted from paraffin-embedded tumor tissue can be used, but also cell-free DNA isolated from liquid biopsies such as blood.In this research project, cfRRBS will be turned into a clinically applicable protocol. A computational analysis pipeline with reference data sets will be built to classify tumors according to their methylation profile. The processing of patient samples and sequencing data will be optimized to achieve a flexible and reliable classifier. The performance of this classifier will then be prospectively validated as a diagnostic tool for CUPs." "Anaesthesia-related allergy: from new insights in pathomechanisms to reliable diagnostic instruments." "Didier Ebo" "Translational Pathophysiological Research (TPR)" "Allergy during anaesthesia is an important health problem with a significant mortality and morbidity. Unfortunately, correct diagnosis is not always straightforward and too frequently overlooked. As a consequence, patients are at unnecessary risk for future life-threatening reactions. Uncertainties associated with skin testing using some of these drugs and the general unavailability of drug-specific immunoglobulin E tests contribute to the frequent failure of adequate investigation and identification of underlying mechanisms, distinguishing IgE-independent from IgE-dependent true anaphylactic reactions in these patients. In this project we would like to explore further the underlying pathomechanisms of immediate type (or IgE-mediated) hypersensitivity reactions that occur during general anaesthesia. For this purpose, we will take opportunity of having developed and validated up-to-date flow cytometric techniques like basophil activation tests and a cellular histamine content assay (HistaFlow®: patented by our laboratory) next to recently developed tools to culture mast cells out of circulating CD34+ progenitor cells. This will enable a combined analysis of surface markers and histamine release after IgE-dependent and IgE-dependent basophil and mast cell activation. Moreover, by having set in place a reference centre with highest throughput of patients in Belgium, it is anticipated that these experiments not only might culminate in the development of novel diagnostic tests to document drug hypersensitivity reactions, but also might help to define save anaesthetic alternatives." "Molecular Epidemiology of tuberculosis at Human-Animal interface in the pastoral areas of Ethiopia with subsequent development of improved diagnostic approach" "Jan Paeshuyse" "Animal and Human Health Engineering (A2H)" "Against the available diagnosis and treatment advances, tuberculosis remains a relentless disease that is one of the leading causes of death from a single infectious agent worldwide. More importantly, the highest burden of tuberculosis is attributed to low- and middle-income countries such as Ethiopia, where limited resources and high population density intersect. Since zoonotic mycobacteria have  numerous host ranges that serve as reservoirs, it can complicate the fight against animal and / or human tuberculosis and have negative consequences and have significant economic problems and threaten livelihoods. Human-to-animal transmission of Mycobacterium tuberculosis complex, including M. Tuberculosis, is well documented in many parts of the world, including Ethiopia, which could have an implication in the epidemiology and control of tuberculosis in human and animal populations. However, livestock farmers and other herdsmen and slaughterhouse workers can also contract the disease via contaminated air droplets via aerosols from infected animals with pulmonary tuberculosis. This is particularly important in Ethiopia, where tuberculosis is endemic, milk pasteurisation is limited and human-animal interaction is unavoidable. Reverse transmission of Mycobacterium tuberculosis complex and other atypical mycobacteria between camel and humans in areas of intense human-animal contact is one of the driving factors for the onset and maintenance of tuberculosis in environments such as camel-herding pastoral areas of Ethiopia. By endorsing the ''Global End TB Strategy'' by 2035, Ethiopia's National End TB Strategy aimed to eliminate tuberculosis epidemics by reducing tuberculosis-related deaths by 95% and by reducing incidental tuberculosis cases by 90% between 2015 and 2035. One of the huge obligations of this programme is to improve access and equitable tuberculosis services for vulnerable and marginalised populations where tuberculosis is concentrated and where most delays occur due to socio-economic and legal barriers. Moreover, research and innovations are also the top priorities that the program has highlighted to sharply bend the country's tuberculosis epidemic curve to meet its ambitious 2035 targets. On the other hand, as part of its livestock master plan, Ethiopia planned to increase its GDP by increasing the production and export of live animals and animal products, and also to meet the growing national demand for affordable animal protein. And camel production is focused on development to meet in the pastoral areas of Ethiopia, where camels are herded primarily as a livelihood and survival strategy of the pastoral communities. However, the sector is not contributing as expected to the national economy and the livelihoods of livestock farmers due to various constraints, diseases such as tuberculosis of which are huge barriers to livestock productivity, exports, food security and public health. Tuberculosis in dromedaries threatens the well-being and livelihoods of pastoral communities, in addition to having a significant economic impact through reduced meat and milk production and creating barriers to international trade in dromedaries and their products. It would therefore be very important to assess the occurrence of Mycobacteria infection at the human-animal interface in the  pastoral communities of camel rearing in Ethiopia, describe the type species and/or strain circulating among humans and camels, and relate the transmission risks while describing the epidemiology and transmission dynamics of Mycobacteria. Such a study is also essential in devising context-appropriate tuberculosis control and prevention interventions. However, the accuracy of diagnostic tests for tuberculosis infection used in animals, mainly camels, is often elusive when diagnosing tuberculosis, and yet there is no reliable diagnostic test for accurate early ante-mortem diagnosis of tuberculosis in dromedaries, indicating the need for exploration and development of a new diagnostic tool. Against this background, this project is designed to study the molecular epidemiology of tuberculosis at the human-animal interface and to assess the bidirectional zoonotic transmission of Mycobacteria species between humans and animals in the dromedary camel-rearing pastoral communities in Ethiopia, and the subsequent development of proof-of-concept for an improved diagnostic approach to animal and human tuberculosis using camelids, nanobodies and phage technology. Key words: Tuberculosis, Mycobacterium, Camel, Pastoral, Ethiopia, Diagnosis, Nanobodies" "Contemporary diagnostic challenges in general internal medicine: focus on updating the approach to fever and inflammation of unknown origin" "Steven Vanderschueren" "Adaptive Immunology, Laboratory for Clinical Infectious and Inflammatory Disorders" "Fever of unknown origin (FUO) and inflammation of unknown origin (IUO) are challenging clinical syndromes characterized by a duration of at least 3 weeks, either fever of 38.3°C (FUO) or elevated inflammatory markers without high-grade fevers (IUO) on several occasions, and no diagnosis despite a baseline workup. The underlying causes are typically categorized as infections, malignancies, noninfectious inflammatory disorders, and miscellaneous disorders. A thoughtful exploration is warranted, and the initial selection of diagnostic tests is based on the presence of potential diagnostic clues, which includes any information obtained from the history, clinical examination, and prior work-up. Earlier studies indicated that potential diagnostic clues may often be misleading, and thus it is important to systematically evaluate their value and limitations. Our research aimed to focus on the diagnostic value of several of these potentially helpful clues. In Chapter 3, we evaluated the importance of the symptom duration and how it alters the diagnostic category distribution. We found that there was little difference between those with symptoms for less than 3 months and those with symptoms for 3 to 12 months. In contrast, those with a symptom duration of more than 12 months were less likely to reach a final diagnosis due to a considerably lower likelihood of infections, malignancies, and miscellaneous disorders. Although the list of underlying causes of FUO/IUO was extensive, we observed only a few infections, malignancies, and miscellaneous disorders among those with long-standing symptoms.  In Chapter 4, we assessed the value of the periodicity of symptoms or symptom pattern and compared those with a recurrent pattern to those with a continuous pattern. A recurrent pattern was observed in one in four patients and was associated only with a lower likelihood of infections at last follow up. However, the recurrent group had considerable diagnostic delay compared to the continuous group, which was most pronounced for the noninfectious inflammatory disorders. Over half of the disorders which were observed at least twice presented with both symptom patterns. In Chapter 5, we compared the diagnostic yield of [18F]-FDG-PET between patients with FUO and IUO as a prior study paradoxically indicated that absence of fever was associated with a higher diagnostic yield. We indeed confirmed that IUO patients more often had a contributive [18F]-FDG-PET compared to those with FUO, which we hypothesized to be related to distinctions in their diagnostic spectrum. In Chapter 6, we further compared the FUO and IUO population, which were assumed to be similar in terms of diagnostic spectrum based on an earlier study, by performing a meta-analysis combining all studies including both FUO and IUO patients. We found that there were no important differences in terms of diagnostic categories apart from infections, which were slightly less likely in those with FUO. We did identify important differences only within the group of noninfectious inflammatory disorders, as IUO patients were less likely to have systemic autoinflammatory disorders and more likely to have vasculitis and rheumatoid arthritis or spondyloarthritis. In Chapter 7, we focused on the specific subgroup of FUO/IUO patients who have no abnormalities on [18F]-FDG-PET during the work-up. Although negative [18F]-FDG-PET imaging may not be considered a typical potential diagnostic clue per se, this particular circumstance is expected to occur more frequently in the future. We found that patients with negative [18F]-FDG-PET imaging had a lower likelihood of reaching a final diagnosis and were less likely to have malignancies. The most important reasons for [18F]-FDG-PET being negative included [18F]-FDG-PET being truly false negative, patients having localized disease for which routine FDG-PET imaging may not be considered appropriate, patients not having any localized disease, and patients not having a disease flare at the time of imaging. Repeat [18F]-FDG-PET was valuable in only a minority. In conclusion, certain clinical scenarios may require a different approach. However, it is important to emphasize that even though the scenarios we evaluated often altered the likelihood of certain diagnostic categories, we were never able to fully exclude a diagnostic category based on these specific characteristics. Optimal management of these diagnostically challenging syndromes will continue to require a thorough knowledge of the disease spectrum and clinical experience." "The performance of saliva specimens for the molecular detection of SARS-CoV-2 and a saliva pooling strategy as a diagnostic tool for epidemiological studies in current SARS-CoV-2 pandemic." "Piet Cools" "Department of Internal Medicine and Pediatrics, Department of Translational Physiology, Infectiology and Public Health, Department of Veterinary Public Health and Food Safety, Department of Diagnostic Sciences" "Nasopharyngeal swabs are recommended for the molecular diagnosis of COVID-19 but offer disadvantages such as dire shortage. Therefore, we aim to validate saliva as an alternative specimen. We further plan to design a saliva pooling strategy as a part of a diagnostic tool for largescale real-time epidemiological testing. Such strategy will be of value in the management of current pandemic."