Deciphering reference intervals and clinical decision limits in equine endocrine diagnostic testing

Reference intervals (RIs) and clinical decision limits (CDLs) are frequently established to facilitate interpretation of values of endocrine biomarkers in the diagnosis of disease. Despite their commonplace use in clinical decision-making, these concepts can be confused. Comparing a test result with a RI provides an estimation as to whether or not the individual is healthy, whereas comparison with a CDL facilitates identification of individuals with a particular disease state or at greater risk of adverse clinical outcomes. In practice, there will also be a range of results for which the discriminative ability of the test is insufficient to inform a specific diagnostic decision. Including a range of uncertain test results, or 'grey zone', between positive and negative avoids the constraint of a binary decision in classifying an individual with a test value above (or below) a single cut-off value as diseased. This review will detail the application of both RIs and CDLs, including defining the range of uncertain test results, in the context of equine endocrinology.