In order to make the most of the information available, the results of BAT can be used in combination with the results of additional checks [15, 17, 44, 45]. histograms showing the manifestation of CD63 and CD203c on the surface of basophils in different conditions. Unstimulated cells (bad control) and cells stimulated with peanut or with anti-IgE (positive control) are displayed. The manifestation of CD63 is Carbamazepine measured as the percentage of positive basophils (quantity of study participants, pollen-food syndrome, not identified, versus, sensitised but tolerant, non-sensitised non-allergic,? em SI? /em activation index Numerous factors may influence the diagnostic overall performance and cut-off ideals of BAT in different studies, some related to the study populace, some related to the study design, some related to the laboratory procedure and to the strategy used for data analysesTable?2. Existing studies are heterogeneous in most of these elements, which limits their comparability and a wider software of the diagnostic cut-offs identified in specific studies. The criterion to diagnose each food allergy is definitely allergen-specific and the diagnostic accuracy may not be the same for different allergens. Additionally, the cut-offs defined in one populace are not necessarily directly transferrable to another populace from a different geographical location assessed inside a different Allergy centre. One limitation of BAT is the fact that a small proportion of individuals tested have non-responder basophils (i.e. basophils that respond to a non-IgE-mediated positive control but not to IgE-mediated stimulants) and therefore have an uninterpretable result for the test. Additional challenges in translating the BAT from a research method to a diagnostic test in the medical center are related to the standardisation of the assay and its reproducibility and also to the cost-effectiveness of including BAT in the diagnostic approach of individuals with suspected food allergy. These elements have not yet been founded and require further study. Table?2 Examples of factors that can influence the diagnostic cut-offs for BAT?in food allergy [4, 14, 39, 40, 41] Study populationPrevalence of the food allergy in the populationOrigin of the study population (e.g. recruited from a specialized Allergy medical center or from the general populace)Geographical locationAssociated respiratory and food allergiesStudy designInclusion criteria (e.g. whether sensitized as well as non-sensitized individuals were included in the study)Gold-standard used like a comparator to determine the diagnostic cut-offsCriteria for carrying out OFC (e.g. whether individuals with Ace a history of anaphylaxis or additional risk factors for any severe reaction or with high levels of IgE or large wheals on pores and skin prick test were included)OFC protocol (e.g. criteria for preventing the OFC, criteria for any positive OFC, intervals between doses and period of OFC)Laboratory procedureInterval between blood collection and the overall performance of BATAllergen components or purified/recombinant allergens usedConcentration of the allergensPre-incubation with IL-3Markers and antibodies (e.g. clones, fluorochromes) used to identify the basophils?and to detect basophil activationFlow cytometry data analysesAdopted gating strategyParameters used as the outcomes of the test [e.g. CD63 or CD203c, % or SI, CD-sens, area under the doseCresponse curve]Definition of bad gateWhether results were corrected for the Carbamazepine backgroundCytometer used and application settings Open in a separate window The strategy adopted to perform the laboratory procedure and to analyse the circulation cytometry data can have a significant impact on the Carbamazepine results acquired for the BAT and, as a result, in its diagnostic accuracy. For example, identifying basophils using an anti-IgE antibody can activate the cells and alter the results obtained having a different method to determine the basophils. The manifestation of particular basophil recognition markers, such as CCR3 [42] and CD123 [43] can change following basophil activation. In a recent study,.