“…Central to the analytical challenges associated with ANA IFA testing are its labor-intensiveness (with significant amounts of training being required for competence), subjectivity in titer and pattern recognition, poor standardization of reagents, and a declining workforce in clinical laboratories (1-4, 15, 30, 32). Thus, in some laboratories, the ANA IFA technique has been replaced by high-throughput, less subjective methods, including enzyme-linked immunosorbent assays (ELISAs) and multiplex assays such as line immunoassays (LIAs) and multiplexed bead assays (MBAs) (21)(22)(23)(24)(25)(26)(27)(28)(29). The non-IFA assays differ in the source, purity, concentration, and binding capacity of the antigens, the reference materials or standards used in assay development, the secondary antibodies (conjugates), and the detection signals.…”