HLA alleles and number of non-self eplets for each patient are shown in Table 1 and Table 2. The remaining eplets (non-self eplets) were then counted and categorized either as reactive or non-reactive based on the cutoff value of the median fluorescence intensity (MFI) value (herein calculated as 500). Non-reactive eplets (assigned blue) were those appearing

in HLA alleles of the panel, which had an MFI value lower than the cutoff value. In contrast, reactive eplets (assigned black) were those appearing only in HLA alleles which had an MFI value higher than the cutoff value. Overall, eplets categorized in this way were used for the classification of the HLA alleles into AMMs and UMMs. Users considered all non-self HLA molecules composed of non-reactive eplets and self-eplets as AMMs. The next step was to compare the results of the conventional Roxadustat chemical structure and automated approaches. Just one eplet, with the same color, should fill correspondent positions in both results.

check details When this rule is broken, there is a disagreement in eplet categorization. A supportive program was created to identify the number of these eplet disagreements between the conventional and automated analyses for each CSV file. It filtered all of the agreeing eplets and showed the number of eplets, AMMs and disagreements in those variables. When disagreements were found, the instructor was invited to critically review the case in order to define whether the error leading to disagreement occurred in the analysis of the single antigen results performed by the conventional or automated method. The four major perceivable features (functionality, reliability, usability and efficiency) were tested to evaluate the quality

of the EpHLA software. Functionality reflects the accuracy in accomplishing the tasks for which the software was designed. Reliability refers to the lack of failures in the software. Usability is an expression of use adequacy as the software must be adequate to the type of user for which it was designed. Thus, it is important that the user can easily understand the concept and application of the program and can learn how to use, operate, and control the tool. Efficiency expresses the capacity of the software to obtain results quickly while using few computer resources. Thymidine kinase Differences in the time spent for the achievement of results using conventional and automated HLAMatchmaker analysis was measured using Student’s t-test and the Mann–Whitney non-parametric test. The disagreements analysis of the numbers of eplets and AMMs among the users were evaluated using the likelihood ratio test after Poisson distribution (H0; lambda < 0.1 vs. H1; lambda ≥ 0.1). The significance levels for all of the tests were established at p < 0.05. The non-experienced group required 60 training hours to be able to analyze single antigen results with HLAMatchmaker using Microsoft Excel format.