Investigation of the dual cascade algorithm in the diagnosis of antinuclear antibodies

Abstract

Abstract Background The dual cascade algorithm which involves screening and confirmation of antinuclear antibodies (ANAs) by further reflex testing is widely used in the detection of ANAs. We aimed to investigate this algorithm which is commonly used in many laboratories. Methods A total of 475 sera obtained from patients with a clinical suspicion of systemic autoimmune rheumatic diseases (SARDs) upon which three expert assessors agreed for interpretation in the indirect immunofluorescence (IIF) test were determined and tested by the line immunoassay (LIA) containing 16 antigens. The results of the tests were statistically compared and evaluated. Results In 141 of the sera (29.7%), there was an agreement between ANA-IIF(+) and LIA(+) results. The overall agreement rate between the two tests for positivity and negativity only was 85.5% with a Cohen’s κ coefficient of 0.69. In 118 of these 141 sera (83.7%), pattern and associated ANA agreement was detected with an overall agreement rate of 80.6% and a Cohen’s κ coefficient of 0.57. The highest agreement between the pattern and associated ANAs was seen in centromere, dense fine speckled (DFS) and cytoplasmic reticular patterns. In these patterns, the rate of anti-centromere-associated protein B (CENP-B), anti-DFS and anti-antimitochondrial antibody M2 (anti-AMA-M2) antibodies were 93.4%, 92.3% and 66.7%, respectively. Conclusions We found an overall moderate agreement between IIF screening and LIA confirmation tests. However, the level of agreement varies according to the pattern type. The discrepancy in agreement rates may cause false reflex test requests. Our results highlight the need for collaboration between clinical and laboratory professionals in selected cases instead of the reflex testing approach.